Corn event MON 87411

ABSTRACT

The invention provides corn event MON 87411, and plants, plant cells, seeds, plant parts, and commodity products comprising event MON 87411. The invention also provides polynucleotides specific for event MON 87411 and plants, plant cells, seeds, plant parts, and commodity products comprising polynucleotides specific for event MON 87411. The invention also provides methods related to event MON 87411.

REFERENCE TO RELATED APPLICATIONS

This application is a divisional of co-pending U.S. application Ser. No.16/404,513, filed May 6, 2019 (pending), which application is adivisional of U.S. application Ser. No. 15/222,789, filed Jul. 28, 2016(now U.S. Pat. No. 10,316,330, which application is a divisional of U.S.application Ser. No. 13/890,027, filed May 8, 2013 (now U.S. Pat. No.9,441,240), which application claims the benefit of U.S. provisionalapplication No. 61/644,368, filed May 8, 2012, the disclosures of whichare herein incorporated by reference in their entirety.

INCORPORATION OF SEQUENCE LISTING

The sequence listing contained in the file named “MONS308US_ST25.txt”,which is 230 kilobytes (size as measured in Microsoft Windows®) and wascreated on May 6, 2013, is filed herewith by electronic submission andis incorporated by reference herein.

FIELD OF THE INVENTION

The invention relates to transgenic Zea mays event MON 87411. The eventprovides dual modes of action for resistance to corn rootworminfestations and tolerance to the herbicide glyphosate. The inventionalso relates to plants, plant parts, plant seeds, plant cells,agricultural products, and methods related to event MON 87411 andprovides nucleotide molecules that are unique to the event and werecreated in connection with the insertion of transgenic DNA into thegenome of a Zea mays plant.

BACKGROUND OF THE INVENTION

Corn (Zea mays) is an important crop in many areas of the world, and themethods of biotechnology have been applied to this crop in order toproduce corn with desirable traits. The expression of an insectresistance or herbicide tolerance transgene in a plant can confer thedesirable traits of insect resistance and/or herbicide tolerance on theplant, but expression of such transgenes may be influenced by manydifferent factors including the orientation and composition of thecassettes driving expression of the individual genes transferred to theplant chromosome, and the chromosomal location and the genomic result ofthe transgene insertion. For example, there can be variation in thelevel and pattern of transgene expression among individual events thatare otherwise identical except for the chromosomal insertion site of thetransgene. There may also be undesirable phenotypic or agronomicdifferences between some events. Therefore, it is often necessary toproduce and analyze a large number of individual plant transformationevents in order to select an event having superior properties relativeto the desirable trait and the optimal phenotypic and agriculturalcharacteristics necessary to make it suitable for commercial purposes.Such selection often requires extensive molecular characterization aswell as greenhouse and field trials with many events over multipleyears, in multiple locations, and under a variety of conditions so thata significant amount of agronomic, phenotypic, and molecular data may becollected. The resulting data and observations must then be analyzed byteams of scientists and agronomists with the goal of selecting acommercially suitable event. Once selected, such an event may then beused for introgressing the desirable trait into other geneticbackgrounds using plant breeding methods, and thus producing a number ofdifferent crop varieties that contain the desirable trait and aresuitably adapted to specific local growing conditions.

To make a transgenic plant containing a single transformation event, aportion of a recombinant DNA construct is transferred into the genome ofa corn cell, and the corn cell is subsequently grown into a plant. Acorn cell into which the event is initially transferred is regeneratedto produce the R₀ generation. The R₀ plant and progeny plants from theR₀ plant can be tested for any desired trait(s), but the effectivenessof the event can be impacted by cis and/or trans factors relative to theintegration site in the transformation event. The phenotype conferred bythe event can also be impacted by the size and design of the DNAconstruct, which can vary by the combination of genetic elements in anexpression cassette, number of transgenes, number of expressioncassettes, and configuration of such elements and such cassettes.Identifying an event with desirable traits can be further complicated byfactors such as plant developmental, diurnal, temporal, or spatialpatterns of transgene expression; or by extrinsic factors, e.g.,environmental plant growth conditions, water availability, nitrogenavailability, heat, or stress. Thus, the ability to obtain an eventconferring a desirable set of phenotypic traits is not readilypredictable.

SUMMARY OF THE INVENTION

The inventors have identified a transgenic corn event MON 87411exhibiting superior properties and performance compared to existingtransgenic corn plants and to new events constructed in parallel. Thecorn event MON 87411 contains three linked expression cassettes whichcollectively confer the traits of corn rootworm resistance andglyphosate herbicide tolerance to corn cells, corn tissues, corn seedand corn plants containing the transgenic event MON 87411. The cornevent MON 87411 provides two modes of action against corn rootworm pestspecies (including Diabrotica spp., especially when the pest isDiabrotica virgifera virgifera (Western Corn Rootworm, WCR), Diabroticabarberi (Northern Corn Rootworm, NCR), Diabrotica virgifera zeae(Mexican Corn Rootworm, MCR), Diabrotica balteata (Brazilian CornRootworm (BZR) or Brazilian Corn Rootworm complex (BCR) consisting ofDiabrotica viridula and Diabrotica speciosa), or Diabroticaundecimpunctata howardii (Southern Corn Rootworm, SCR)). Dual modes ofaction provide redundancy and reduces significantly the likelihood ofthe development of resistance to the pest control traits.

The event MON 87411 is characterized by specific unique DNA segmentsthat are useful in detecting the presence of the event in a sample. Asample is intended to refer to a composition that is eithersubstantially pure corn DNA or a composition that contains corn DNA. Ineither case, the sample is a biological sample, i.e., it containsbiological materials, including but not limited to DNA obtained orderived from, either directly or indirectly, from the genome of cornevent MON 87411. “Directly” refers to the ability of the skilled artisanto directly obtain DNA from the corn genome by fracturing corn cells (orby obtaining samples of corn that contain fractured corn cells) andexposing the genome DNA for the purposes of detection. “Indirectly”refers to the ability of the skilled artisan to obtain the target orspecific reference DNA, i.e. a novel and unique junction segmentdescribed herein as being diagnostic for the presence of the event MON87411 in a particular sample, by means other than by direct viafracturing of corn cells or obtaining a sample of corn that containsfractured corn cells. Such indirect means include but are not limited toamplification of a DNA segment that contains the DNA sequence targetedby a particular probe designed to bind with specificity to the targetsequence, or amplification of a DNA segment that can be measured andcharacterized, i.e. measured by separation from other segments of DNAthrough some efficient matrix such as an agarose or acrylamide gel orthe like, or characterized by direct sequence analysis of the ampliconor cloning of the amplicon into a vector and direct sequencing of theinserted amplicon present within such vector. Alternatively, a segmentof DNA corresponding to the position within the corn chromosome at whichthe transgenic DNA was inserted into the corn chromosome and which canbe used to define the event MON 87411, can be cloned by various meansand then identified and characterized for its presence in a particularsample or in a particular corn genome. Such DNA segments are referred toas junction segments or sequences, and can be any length of inserted DNAand adjacent (flanking) corn chromosome DNA so long as the point ofjoining between the inserted DNA and the corn genome is included in thesegment. SEQ ID NO:12 and SEQ ID NO:21 and the reverse complement ofeach of these are representative of such segments.

The specific sequences identified herein may be present uniquely inevent MON 87411, or the construct comprised therein, and theidentification of these sequences, whether by direct sequence analysis,by detecting probes bound to such sequences, or by observing the sizeand perhaps the composition of particular amplicons described herein,when present in a particular corn germplasm or genome and/or present ina particular biological sample containing corn DNA, are diagnostic forthe presence of the event MON 87411, or the construct comprised therein,in such sample. It is known that the flanking genomic segments (i.e.,the corn genome segments of DNA sequence adjacent to the insertedtransgenic DNA) are subject to slight variability and as such, thelimitation of at least 99% or greater identity is with reference to suchanomalies or polymorphisms from corn genome to corn genome. Nucleotidesegments that are completely complementary across their length incomparison to the particular diagnostic sequences referenced herein areintended to be within the scope of the present invention.

The position of the nucleotide segments of the present inventionrelative to each other and within the corn genome are illustrated inFIG. 3 and the nucleotide sequence of each is illustrated as set forthin SEQ ID NO:1. Nucleotide segments that characterize the event MON87411 and which are diagnostic for the presence of event MON 87411, orthe construct comprised therein, in a sample include SEQ ID NO:1, SEQ IDNO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7,SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, SEQ ID NO:12, SEQ ID NO:14, SEQID NO:16, SEQ ID NO:18, SEQ ID NO:19, SEQ ID NO:20, SEQ ID NO:21, SEQ IDNO:22, SEQ ID NO:23, SEQ ID NO:24, and SEQ ID NO:25; SEQ ID NO:41, SEQID NO:42, SEQ ID NO:43, SEQ ID NO:44, SEQ ID NO:45, SEQ ID NO:49, SEQ IDNO:50, SEQ ID NO:51, or SEQ ID NO:52. These presence of one, or two, ormore of these nucleotide sequences in a sample, when such samplecontains corn tissue and thus corn DNA, are diagnostic for the presenceof the event MON 87411, or the construct comprised therein.

It is intended by use of the word “derived”, that a particular DNAmolecule is in the corn plant genome, or is capable of being detected incorn plant DNA. “Capable of being detected” refers to the ability of aparticular DNA segment to be amplified and its size and or sequencecharacterized or elucidated by DNA sequence analysis, and can also referto the ability of a probe to bind specifically to the particular DNAsegment, i.e. the target DNA segment, and the subsequent ability todetect the binding of the probe to the target. The particular DNAsegment or target DNA segment of the present invention is present withincorn that contains the insertion event MON 87411.

By reference to corn it is intended that corn cells, corn seed, cornplant parts and corn plants are within the scope of the presentinvention so long as each embodiment contains a detectable amount of DNAcorresponding to any one, two, or more of the segments that aredescribed herein as being diagnostic for the presence of the corn eventMON 87411 DNA. Corn plant parts include cells; pollen; ovules pods;flowers and flower parts such as the cob, silk, and tassel; root tissue;stem tissue; and leaf tissue. Commodity products that are made from cornin which a detectable amount of the segments of DNA described herein asbeing diagnostic for the presence of the event MON 87411 are within thescope of the invention. Such commodity products may include whole orprocessed corn seeds, animal feed containing corn or corn by-products,corn oil, corn meal, corn flour, corn starch, corn flakes, corn bran,corn biomass and stover, and fuel products and fuel by-products whenmade from corn or corn plants and plant parts.

The DNA of corn event MON 87411 is typically present in each cell and ineach chromosome of the corn plant, corn seed, and corn tissuescontaining the event. As the corn genome is transmitted to progeny inMendelian fashion, if a corn plant were homozygous, each progeny cornplant and cell would contain the event DNA on each of the parentalchromosomes generated to the progeny from the parent(s). However, if thecorn genome containing the event MON 87411 DNA is a heterozygous orhybrid parent, then only fifty percent of the pollen and fifty percentof the ovules engaged in mating from hybrid parents will contain thecorn event MON 87411 DNA, resulting in a mixed population of progenythat contain the event MON 87411 DNA, and the percentage of such progenyarising from such crosses with hybrids can range anywhere from aboutfifty to about seventy five percent having the event MON 87411 DNAtransmitted to such progeny.

The DNA molecules of the present invention may be unique to the cornevent MON 87411 inserted DNA or the two junctions between the transgenicinserted DNA and the corn genome DNA that is adjacent to either end ofthe inserted DNA. These molecules, when present in a particular sampleanalyzed by the methods described herein using the probes, primers andin some cases using DNA sequence analysis, may be diagnostic for thepresence of an amount of event MON 87411 corn in that sample. Such DNAmolecules unique to the corn event MON 87411 DNA can be identified andcharacterized in a number of ways, including by use of probe nucleicacid molecules designed to bind specifically to the unique DNA moleculesfollowed by detection of the binding of such probes to the unique DNA,and by thermal amplification methods that use at least two different DNAmolecules that act as probes but the sequence of such molecules may besomewhat less specific than the probes described above. The skilledartisan understands that contacting a particular target DNA with a probeor primer under appropriate hybridization conditions will result in thebinding of the probe or primer to the targeted DNA segment.

The DNA molecules of the present invention that are target segments ofDNA are capable of amplification and, when detected as one or moreamplicons of the represented length obtained by amplification methods ofa particular sample, may be diagnostic for the presence of event MON87411, or the construct comprised therein, in such sample. Such DNAmolecules or polynucleotide segments have the nucleotide sequences asset forth in each of, SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ IDNO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9,SEQ ID NO:10, SEQ ID NO:12, SEQ ID NO:14, SEQ ID NO:16, SEQ ID NO:21,SEQ ID NO:25, SEQ ID NO:41, SEQ ID NO:42, SEQ ID NO:43, SEQ ID NO:44,SEQ ID NO:45, SEQ ID NO:49, SEQ ID NO:50, SEQ ID NO:51, and SEQ IDNO:52, and are further defined herein and in the examples below. Primermolecules and/or probes may be provided in kit form along with thenecessary reagents, including controls, and packaged together withinstructions for use.

Recombinant DNA molecules of the present invention are deemed to bewithin the scope of the present invention when present within or derivedfrom a microorganism. A microorganism is intended to include anymicroscopic cell, whether prokaryote or eukaryote or otherwise thatcontains DNA within a genome or chromosome or an extra-chromosomal DNAstructure more commonly referred to as a plasmid or vector. Microscopicorganisms include bacteria (prokaryotes) and cells corresponding tohigher life forms (eukaryotes) which are beneath the visual range of theaverage human, typically beneath fifty cubic microns and more generallybeneath ten cubic microns. Bacteria are common microscopicmicroorganisms that more likely than not could contain a vector orplasmid that contains one or more or all of the novel DNA segments ofthe present invention, including each of the respective expressioncassettes present as set forth in SEQ ID NO: 1. Plant cells andparticularly corn plant cells are within the scope of the invention whenthese contain any one, two, or more or all of the novel DNA segments ofthe present invention.

Probes for use herein are typically characterized as DNA molecules orpolynucleotide segments of sufficient length to function under stringenthybridization conditions as defined herein to bind with a particulartarget DNA segment, i.e., a unique segment of DNA present within anddiagnostic for the presence of, event MON 87741 DNA in a sample. Such aprobe can be designed to bind only to a single junction or other novelsequence present only in the corn event MON 87411 DNA, or to two or moresuch single junction segments. In any event, the detection of thebinding of such a probe to a DNA molecule in a particular samplesuspected of containing corn DNA is diagnostic for the presence of cornevent MON 87411 in the sample.

Primers are typically provided as pairs of different oligonucleotides orpolynucleotide segments for use in a thermal amplification reactionwhich amplifies a particular DNA target segment. Each primer in the pairis designed to bind to a rather specific segment of DNA within or nearto a segment of DNA of interest for amplification. The primers bind insuch way that these then act as localized regions of nucleic acidsequence polymerization resulting in the production of one or moreamplicons (amplified target segments of DNA). In the present invention,use of primers designed to bind to unique segments of corn event MON87411 DNA in a particular biological sample and that amplify particularamplicons containing one or more of the junction segments describedherein, and the detection and or characterization of such amplicons uponcompletion or termination of the polymerase reaction, is diagnostic forthe presence of the corn event MON 87411 in the particular sample. Theskilled artisan is well familiar with this amplification method and norecitation of the specifics of amplification is necessary here.

Corn plants, corn plant cells, corn plant tissues and corn seed areinsensitive to glyphosate herbicide applications due to expression of aglyphosate insensitive CP4 EPSPS enzyme from a rice Rcc3 promoter in anexpression cassette at the 3′ distal end as set forth in SEQ ID NO: 1.Such seed may be sown into a field. Several days after germination andthe appearance of shoots, a weed controlling effective amount ofglyphosate herbicide may be applied, which will eliminate substantiallyall of the weeds in the field but will allow for the continued growthand development of corn plants containing the corn event MON 87411 DNA.The plants are also resistant to infestation by corn rootworms of allknown species of rootworm Diabrotica, including but not limited toDiabrotica virgifera virgifera (Western Corn Rootworm, WCR), Diabroticabarberi (Northern Corn Rootworm, NCR), Diabrotica virgifera zeae(Mexican Corn Rootworm, MCR), Diabrotica balteata (Brazilian CornRootworm (BZR) or Brazilian Corn Rootworm complex (BCR) consisting ofDiabrotica viridula and Diabrotica speciosa), and Diabroticaundecimpunctata howardii (Southern Corn Rootworm, SCR). The resistanceto Diabrotica species arises in connection with the expression of twodifferent DNA segments that are operably and covalently linked withinthe inserted transgenic DNA: a dsRNA is transcribed from the expressioncassette at the 5′ proximal end of the inserted transgenic DNA as setforth in SEQ ID NO:1 and as illustrated in FIG. 1 by the position of [G]SEQ ID NO:12, and targets for suppression an essential gene in cornrootworms; and a coleopteran toxic Cry3Bb protein is expressed from anexpression cassette (approximately centered in SEQ ID NO:1 as shown inFIG. 1 by the position of [H] SEQ ID NO: 14) centered between thecassette expressing dsRNA [G] and the cassette at the 3′ distal end ofthe inserted transgenic DNA as set forth in SEQ ID NO:1 (a glyphosatetolerance expression cassette illustrated in FIG. 1 by [I] SEQ IDNO:16). The dsRNA targets for suppression a yeast orthologous genereferred to as snf7 and is expressed from a CAMV e35S promoter, whilethe Cry3Bb protein is expressed from a Zea mays PIIG promoter. The dsRNAand the Cry3Bb protein are agents toxic to corn rootworm species.

The promoters driving expression of the dsRNA and Cry3Bb toxic agentsare divergently positioned so that expression from each promoter of therespective toxic agent is away from a point centered between the twopromoters, i.e., transcription of each expression cassette proceeds inopposite directions and does not converge. The glyphosate tolerance CP4EPSPS expression cassette is downstream of, i.e. proximal to the 3′ endas set forth in SEQ ID NO:1 and 3′ distal to the cassette drivingexpression of the Cry3Bb protein. The cassettes driving expression ofCry3Bb and EPSPS produce their respective proteins using a tandemorientation of transcription, Cry3Bb upstream of the EPSPS, andtranscribed in the same orientation, but each from their separaterespective promoters. Leaving the dsRNA expression cassette and theglyphosate tolerance cassette intact and positioned at the distal endsof the DNA segment intended for insertion into the corn genome, othervariant constructs were produced in which the orientation of the Cry3Bbcassette was inverted or reversed relative to the design present in theevent MON 87411 DNA. These variant constructs utilized the Zea mays PIIGpromoter or a rice Rcc3 promoter to drive expression of Cry3Bb.

Transgenic events containing only these variant constructs/orientationsof the Cry3Bb expression cassette were compared to the event MON 87411and to the currently available commercial events MON863 (containing onlya Cry3Bb expression cassette), MON88017 (containing a Cry3Bb expressioncassette operably linked to a CP4 EPSPS expression cassette), andDAS-59122-7 (containing three operably linked expression cassettes, twoexpressing in tandem the dual Bt toxin components Cry34 and Cry35 alongwith a gene conferring glufosinate tolerance). The results asillustrated below in the examples show that the event MON 87411exhibited superior properties for root directed expression of the Cry3Bbprotein and the plurality of transgenic events produced using theconstruct used for generating the event MON 87411 were each more likelythan other events produced with other constructs to exhibit efficaciouscontrol of corn rootworms.

Corn plants of the present invention and parts thereof including seed,each containing the DNA corresponding to event MON 87411, are within thescope of the present invention. Such plants are resistant to cornrootworm infestation and are insensitive to applications of theherbicide glyphosate. Such plants include hybrids containing only oneMON 87411 allele, i.e., a genome characterized as heterozygous withreference to the locus corresponding to the event MON 87411 DNA. Suchhybrids are produced by breeding with desirable germplasm to insurehybrid vigor and other agriculturally desirable properties of corn.Hybrids may be produced by any number of methods but a preferred methodtakes advantage of a first inbred (homozygous) parent that contains theevent MON 87411 specific allele on both chromosomes at the locus atwhich the event MON 87411 DNA is inserted, and breeding the first inbredtogether with a second inbred which does not contain the MON 87411 DNA.Both parental inbred varieties will have one or more advantageousproperties desirable in the progeny seed, i.e. the hybrid seed.

A transgenic property or allele conferring some additional trait to aplant containing the event MON 87411 DNA is particularly desirable. Suchtransgenic alleles include other transgenic events conferring cornrootworm resistance, including but not limited to events such asDAS-59122-7; MIR604; and 5307. Each of these events provides asupplemental corn rootworm toxic agent (DAS-59122-7 provides PS149B1(Cry34/Cry35) exhibiting rootworm toxic properties and herbicidetolerance to glufosinate; MIR604 provides a modified Cry3Aa exhibitingrootworm toxic properties; event 5307 provides FR8a gene exhibitingrootworm toxic properties). Providing additional corn rootwormresistance traits such as these may decrease the likelihood of thedevelopment of resistance to any one of the corn rootworm toxic agentsprovided. Other desirable traits include yield and stress resistance ortolerance traits, nitrogen fixation traits, traits modulating the use ofwater, resistance to fungal infestation, resistance to herbicides suchas dicamba (MON 87427), glufosinate, and the like, as well as resistanceto lepidopteran infestations. Lepidopteran infestation resistance traitshave been provided in the art and include the transgenic corn events(and respective lepidopteran active proteins) MON810 (Cry1Ab), MON 89034(Cry1A.105 and Cry2Ab); TC1507 (Cry1Ac and Cry1Fa); DAS-06275-8 alsoknown as TC-6275 (Cry1Fa and bar (providing glufosinate tolerance));MIR162 (Vip3Aa), BT176 (Cry1Ab); and BT11 (Cry1Ab).

An alternative to providing any combination or all of these traits in asingle plant, particularly the insect resistance traits corresponding tothe event MON 87411 traits, the other listed corn rootworm resistancetraits, or the lepidopteran resistance traits, would be to provide thesein various combinations of seed blends, in which certain seed in theblend contain the MON 87411 traits and some combination of only thelisted coleopteran resistance traits and act together below the groundto prevent infestations of corn rootworms, while other seed in the blendcontain only the lepidopteran resistance traits and confer resistance tolepidopteran infestations of corn above the ground. In this way, theseed in the blend provide refuge for each other, i.e. the coleopteranprotected seed and plants act as a refuge for the plants conferringlepidopteran resistance, and vice versa. Typically however, these traitswould be provided in some trait combination or package in which the MON87411 traits would be provided together in a single plant by breedingwith one or more of the lepidopteran resistance traits to provide acomplete package of pest resistance to the crop in the field, and asmall percentage of the seed (perhaps between 1 and 20 percent or anynumber in between including 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14,15, 16, 17, 18, or 19 percent) would be traited only for herbicidetolerance and would lack any pest protection traits and would be plantedinto the field in a mix randomly with the pest resistance traited seedor as a structured (separate) stand of crops would act as a refuge bothfor the pests that attack corn plants above the ground and pests thatattack corn plants below the ground.

In one aspect, the invention therefore provides a method of protecting afield of corn plants comprising cultivating a field of corn plantscomprised of from about 50 to about 100 percent of corn plantscomprising corn event MON 87411.

The construct inserted into the event MON 87411 provides particularadvantages relative to the EPSPS expression cassette. First, thepresence of this cassette provides for ease of selection of thetransgenic events into which the construct has been inserted. Second,the cassette provides for control of weeds in a field into which seedcorresponding to event MON 87411 have been planted. The field containingsuch MON 87411 plants can be sprayed with an effective amount ofglyphosate to control the growth of weeks in the field that aresusceptible to glyphosate. For weeds that are not susceptible toglyphosate. As noted above, other transgenic events that provide fortolerance to other herbicides such as to dicamba or to glufosinate canbe bred into a single hybrid along with the event MON 87411, thusproviding an efficient means for controlling weeds in a field byapplying two or more of the herbicides glyphosate, dicamba, orglufosinate, as the likelihood that weeds would be present that exhibittolerance to two or more of these herbicides would be unlikely, and insuch case, the corn crop would consist of hybrids that exhibitresistance to such applications of herbicide combinations.

In one aspect, the invention provides a DNA molecule comprising (a) therecombinant polynucleotide as set forth in SEQ ID NO:12; and (b) therecombinant polynucleotide as set forth in SEQ ID NO:14; and (c) therecombinant polynucleotide as set forth in SEQ ID NO:16, wherein saidrecombinant polynucleotide sequences are linked together byphosphodiester linkage. In one embodiment, the DNA molecule comprisesSEQ ID NO:4.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 is a diagrammatical representation of the transgenic insert inthe genome of corn event MON 87411: [A] represents SEQ ID NO:1, which isthe contiguous sequence of the transgenic DNA insert integrated into thegenome of corn LH244 and 5′ and 3′ genomic DNA flanking the insertedDNA; [B] and [C] correspond to the relative positions of SEQ ID NOs:2and 3, which form the 5′ and 3′ transgene/genomic DNA junction sequencesof event MON 87411, respectively; [D] represents SEQ ID NO:4, which isthe sequence of the transgenic DNA insert integrated into the genomeresulting in event MON 87411; [E] corresponds to the relative positionsof SEQ ID NO:5, SEQ ID NO:6 and SEQ ID NO:7, each spanning the 5′junction between the terminal ends of the transgenic inserted DNA andthe flanking genomic DNA; [F] corresponds to the relative positions ofSEQ ID NO:8, SEQ ID NO:9 and SEQ ID NO:10, each spanning the 3′ junctionbetween the terminal ends of the transgenic inserted DNA and theflanking genomic DNA; [G], [H] and [I] respectively represent the threedifferent expression cassettes corresponding to the transgenic DNAconstruct inserted into the corn plant genome resulting in event MON87411; [J], and [K] represent oligonucleotide primers, oligonucleotideprobes, and DNA amplicons corresponding to the junctions of event MON87411.

FIG. 2 illustrates eleven different DNA constructs, (417, 416, 418, 419,402, 403, 404, 423, 405, 406, and 890) engineered to express up to threedistinct cassettes, including two plant-incorporated protectant (PIP)cassettes, targeting Western corn rootworm (WCR), and a single herbicidetolerance cassette. The two PIP cassettes include (a) an expressioncassette for a Dv_Snf7o 240-mer inverted repeat, and (b) an expressioncassette for a Cry3Bb protein. Each of the constructs depicted comprisethese expression cassettes in varying order and orientation. Constructs405 and 406 contain no herbicide tolerance cassette and construct 890comprises only a single expression cassette for a Dv_Snf7o 240-merinverted repeat. The three constructs comprise a total of sixteengenetic elements from the Left Border (LB) through to the Right Border(RB): [1] LB; [2] Ps.RbcS2-E9 3′ UTR; [3] 240-mer Dv_Snf7o invertedrepeat gene; [4] Corn DnaK intron; [5] CaMV 35S leader; [6] eCaMV 35Spromoter; [7] Corn PIIG promoter; [8] Wheat Lhcb1 leader; [9] Rice Act1intron; [10] cry3Bb ORF; [11] Wheat Hsp17 3′ UTR; [12] Rice TubA(promoter, leader, intron); [13] CTP; [14] CP4 EPSPS; [15] Rice TubA 3′UTR; and [16] RB.

FIG. 3 [A]-[N] and [aa]-[mm] illustrate the operably linked elements andflanking corn genome and their position relative to each other as theseare presented within the transgenic DNA insertion position in the cornevent MON87411 genome. The following descriptions identify thecomposition, function and position for each of the elements as set forthin SEQ ID NO:1.

[A] nucleotide position 1-500 as set forth in SEQ ID NO:1 corresponds tocorn genome DNA adjacent to the transgenic inserted DNA in corn eventMON87411, which in this case is arbitrarily assigned as the 5′ end ofthe transgenic inserted DNA.

[B] nucleotide position 807-1439 as set forth in SEQ ID NO:1 correspondsto the reverse complement sequence of a Pisum sativum ribulose bisphosphate carboxylase small subunit E9 3′ transcription termination andpolyadenylation signal.

[C] nucleotide position 1469-2098 as set forth in SEQ ID NO:1corresponds to the reverse complement sequence designed to be expressedas an RNA molecule that folds into a 240 nucleotide dsRNA and 150nucleotide hairpin structure that is designed to target for suppressionthe Diabrotica species orthologue of a yeast gene encoding an Snf7protein when provided in the diet of a Diabrotica species. A first 240nucleotide segment corresponding to a portion of the Diabrotica snf7orthologous gene is provided at nucleotide position 1469-1708 as setforth in SEQ ID NO:1, a second 240 nucleotide segment corresponding tothe reverse complement of the first segment is set forth at nucleotideposition 1850-2098 as set forth in SEQ ID NO:1, and the first and thesecond segments are operably linked by a 150 nucleotide spacer atnucleotide position 1709-1858 as set forth in SEQ ID NO:1.

[D] nucleotide position 2135-2938 as set forth in SEQ ID NO:1corresponds to the reverse complement sequence of an intron derived froma Zea mays dnaK gene.

[E] nucleotide position 2839-3298 as set forth in SEQ ID NO:1corresponds to the reverse complement of a Cauliflower mosaic virusenhanced 35S promoter sequence and an untranslated 5′ leader sequence.This promoter, the associated untranslated leader, the intron element[D] and the transcription termination and polyadenylation element [B]regulate the expression of element [C] in corn plant cells.

[F] nucleotide position 3586-4534 as set forth in SEQ ID NO:1corresponds to a promoter sequence derived from a Zea mays physicalimpedance induced protein gene (Zm.PIIG). This promoter, the associateduntranslated leader [G], the intron element [H] and the transcriptiontermination and polyadenylation element [J] regulate the expression ofelement [I]. This promoter is oriented relative to the promoter [E] suchthat each promoter ([E] and [F]) will drive divergent expression oftheir respective elements ([C] and [I]) (see block arrows in FIG. 2where the arrows are representative of the respective promoters ([E] and[F]) in the indicated direction of expression from the respectivepromoter).

[G] nucleotide position 4541-4601 as set forth in SEQ ID NO:1corresponds to an untranslated 5′ leader sequence derived from aTriticum aestivum light harvesting complex b1 gene (Ta.Lhcb1).

[H] nucleotide position 4618-5097 as set forth in SEQ ID NO:1corresponds to an intron sequence derived from an Oryza sativa Actin-1gene (Os.Act1).

[I] nucleotide position 5107-7068 as set forth in SEQ ID NO:1corresponds to the nucleotide sequence encoding a Cry3Bb corn rootwormtoxic protein (cry3Bb). The encoded Cry3Bb protein is pesticidal whenprovided in the diet of a Diabrotica (corn rootworm) species.

[J] nucleotide position 7088-7297 as set forth in SEQ ID NO:1corresponds to the sequence of a Triticum aestivum heat shock protein 17(HSP17) transcription termination and polyadenylation signal.

[K] nucleotide position 7346-9526 as set forth in SEQ ID NO:1corresponds to a contiguous promoter-leader-intron sequence derived froman Oryza sativa alpha tubulin-3 gene (TubA-3). This promoter, with theassociated leader and intron, and the transcription termination andpolyadenylation element [M] regulate the expression of element [L].

[L] nucleotide position 9531-11126 as set forth in SEQ ID NO:1corresponds to sequence of an Arabidopsis thaliana cytoplasmic targetingpeptide (CTP; from nucleotide position 9531-9758), and a sequence of anEPSPS derived from Agrobacterium CP4 (from nucleotide position9759-11126). When this sequence is transcribed and translated intoprotein in a corn plant cell, the CTP is operably linked to the EPSPS.When expressed in corn plant cells comprising event MON87411, thisCTP-EPSPS provides tolerance to the herbicide glyphosate.

[M] nucleotide position 11134-11715 as set forth in SEQ ID NO:1corresponds to the sequence of an Oryza sativa alpha tubulin-3 gene(TubA-3) transcription termination and polyadenylation signal.

[N] nucleotide position 11749-12248 as set forth in SEQ ID NO:1corresponds to corn genome DNA adjacent to the transgenic inserted DNAin corn event MON87411, which in this case is arbitrarily assigned asthe 3′ end of the transgenic inserted DNA.

[aa] nucleotide position 501-806 as set forth in SEQ ID NO:1 correspondsto the portion of the Agrobacterium tumefaciens octopine left bordersequence of the 417 construct adjacent to the genome at the arbitrarilyassigned 5′ end of the transgenic DNA inserted into the corn genome toform event MON 87411. The 5′ end of [aa] as set forth in SEQ ID NO: 1 islinked to the 3′ end of element [A] to form the unique 5′ transgenicinserted DNA/corn genome junction encompassed by SEQ ID NO:5, SEQ IDNO:6, SEQ ID NO:7, and SEQ ID NO:21. The 3′ end of element [aa] islinked to the 5′ end of element [B] to form a unique junction within thetransgenic inserted DNA that is encompassed by SEQ ID NO:41.

[bb] nucleotide position 1440-1468 as set forth in SEQ ID NO:1corresponds to the an intervening sequence between elements [B] and [C].The 5′ end of [bb] as set forth in SEQ ID NO:1 is linked to the 3′ endof element [B], and the 3′ end of element [bb] is linked to the 5′ endof element [C] to form a unique junction, encompassed by SEQ ID NO:42,within the transgenic DNA inserted into the corn genome to form eventMON 87411.

[cc] nucleotide position 2099-2134 as set forth in SEQ ID NO:1corresponds to the an intervening sequence between elements [C] and [D].The 5′ end of [cc] as set forth in SEQ ID NO:1 is linked to the 3′ endof element [C], and the 3′ end of element [cc] is linked to the 5′ endof element [D] to form a unique junction, encompassed by SEQ ID NO:43,within the transgenic DNA inserted into the corn genome to form eventMON 87411.

[ee] nucleotide position 3299-3585 as set forth in SEQ ID NO:1corresponds to the an intervening sequence between elements [E] and [F].The 5′ end of [ee] as set forth in SEQ ID NO:1 is linked to the 3′ endof element [E], and the 3′ end of element [ee] is linked to the 5′ endof element [F] to form a unique junction, encompassed by SEQ ID NO:44,within the transgenic DNA inserted into the corn genome to form eventMON 87411.

[ff] nucleotide position 4535-4540 as set forth in SEQ ID NO:1corresponds to the an intervening sequence between elements [F] and [G].The 5′ end of [ff] as set forth in SEQ ID NO:1 is linked to the 3′ endof element [F], and the 3′ end of element [ff] is linked to the 5′ endof element [G] to form a unique junction, encompassed by SEQ ID NO:45,within the transgenic DNA inserted into the corn genome to form eventMON 87411.

[gg] nucleotide position 4602-4617 as set forth in SEQ ID NO:1corresponds to the an intervening sequence between elements [G] and [H].The 5′ end of [gg] as set forth in SEQ ID NO:1 is linked to the 3′ endof element [G], and the 3′ end of element [gg] is linked to the 5′ endof element [H] to form a junction, encompassed by SEQ ID NO:46, withinthe transgenic DNA inserted into the corn genome to form event MON87411, but which is not unique to event MON 87411.

[hh] nucleotide position 5098-5106 as set forth in SEQ ID NO:1corresponds to the an intervening sequence between elements [H] and [I].The 5′ end of [hh] as set forth in SEQ ID NO:1 is linked to the 3′ endof element [H], and the 3′ end of element [hh] is linked to the 5′ endof element [I] to form a junction, encompassed by SEQ ID NO:47, withinthe transgenic DNA inserted into the corn genome to form event MON87411, but which is not unique to event MON 87411.

[ii] nucleotide position 7069-7087 as set forth in SEQ ID NO:1corresponds to the an intervening sequence between elements [I] and [J].The 5′ end of [ii] as set forth in SEQ ID NO:1 is linked to the 3′ endof element [I], and the 3′ end of element [ii] is linked to the 5′ endof element [J] to form a junction, encompassed by SEQ ID NO:48, withinthe transgenic DNA inserted into the corn genome to form event MON87411,but which is not unique to event MON 87411.

[jj] nucleotide position 7298-7345 as set forth in SEQ ID NO:1corresponds to the intervening sequence between elements [J] and [K].The 5′ end of [jj] as set forth in SEQ ID NO:1 is linked to the 3′ endof element [J], and the 3′ end of element [jj] is linked to the 5′ endof element [K] to form a unique junction, encompassed by SEQ ID NO:49,within the transgenic DNA inserted into the corn genome to form eventMON 87411.

[kk] nucleotide position 9527-9530 as set forth in SEQ ID NO:1corresponds to the intervening sequence between elements [K] and [L].The 5′ end of [kk] as set forth in SEQ ID NO:1 is linked to the 3′ endof element [K], and the 3′ end of element [kk] is linked to the 5′ endof element [L] to form a unique junction, encompassed by SEQ ID NO:50,within the transgenic DNA inserted into the corn genome to form eventMON 87411.

[ll] nucleotide position 11127-11133 as set forth in SEQ ID NO:1corresponds to the an intervening sequence between elements [L] and [M].The 5′ end of [ll] as set forth in SEQ ID NO:1 is linked to the 3′ endof element [L], and the 3′ end of element [ll] is linked to the 5′ endof element [M] to form a unique junction, encompassed by SEQ ID NO:51,within the transgenic DNA inserted into the corn genome to form eventMON 87411.

[mm] nucleotide position 11716-11748 as set forth in SEQ ID NO:1corresponds to the a portion of the Agrobacterium tumefaciens nopalineright border sequence of the 417 construct adjacent to the genome at thearbitrarily assigned 3′ end of the transgenic DNA inserted into the corngenome to form event MON 87411. The 5′ end of [mm] as set forth in SEQID NO:1 is linked to the 3′ end of element [M], and the 3′ end ofelement [mm] is linked to the 5′ end of element [N] to form a uniquetransgenic inserted DNA/corn genome junction encompassed by SEQ IDNO:52.

FIG. 4 Illustration of cassette orientation for vectors tested to showhigher efficacy of divergent promoters driving expression of cornrootworm toxic agents compared to vectors with a tandem orientation ofpromoters driving expression of corn rootworm toxic agents.

BRIEF DESCRIPTION OF THE SEQUENCES

SEQ ID NO:1 is a nucleotide sequence of event MON 87411, and representsfrom 5′ to 3′, a segment of the 5′ genomic DNA flanking (adjacent to)the inserted transgenic DNA (500 nucleotides), the inserted transgenicDNA (11,248 nucleotides), and a segment of the 3′ genomic DNA flanking(adjacent to) the inserted transgenic DNA (500 nucleotides) in event MON87411.

SEQ ID NO:2 is a nucleotide junction sequence of event MON 87411, andrepresents from 5′ to 3′, a segment of the 5′ genomic DNA adjacent tothe inserted transgenic DNA (500 nucleotides), and the insertedtransgenic DNA border remnant (263 nucleotides) of event MON 87411.

SEQ ID NO:3 is a nucleotide junction sequence of event MON 87411, andrepresents from 5′ to 3′, the inserted transgenic DNA border remnant (15nucleotides), and a segment of the 3′ genomic DNA adjacent to theinserted genomic DNA (500 nucleotides) of event MON 87411.

SEQ ID NO:4 is a nucleotide sequence of event MON 87411, and representsthe inserted genomic DNA (11248 nucleotides) of event MON 87411.

SEQ ID NO:5 is a nucleotide junction sequence of event MON 87411, andrepresents from 5′ to 3′, a segment of the 5′ genomic DNA adjacent tothe inserted transgenic DNA (50 nucleotides), and the insertedtransgenic DNA border remnant (263 nucleotides) of event MON 87411.

SEQ ID NO:6 is a nucleotide junction sequence of event MON 87411, andrepresents from 5′ to 3′, a segment of the ′5 genomic DNA adjacent tothe inserted transgenic DNA (110 nucleotides), and the insertedtransgenic DNA border remnant (263 nucleotides) of event MON 87411.

SEQ ID NO:7 is a nucleotide junction sequence of event MON 87411, andrepresents from 5′ to 3′, a segment of the 5′ genomic DNA adjacent tothe inserted transgenic DNA (145 nucleotides), and the insertedtransgenic DNA border remnant (263 nucleotides) of event MON 87411.

SEQ ID NO:8 is a nucleotide junction sequence of event MON 87411, andrepresents from 5′ to 3′, a segment of the inserted transgenic DNA (83nucleotides), and a segment of the 3′ genomic DNA adjacent to theinserted transgenic DNA (34 nucleotides) of event MON 87411.

SEQ ID NO:9 is a nucleotide junction sequence of event MON 87411, andrepresents from 5′ to 3′, a segment of the inserted transgenic DNA (83nucleotides), and a segment of the 3′ genomic DNA adjacent to theinserted transgenic DNA (90 nucleotides) of event MON 87411.

SEQ ID NO:10 is a nucleotide junction sequence of event MON 87411, andrepresents from 5′ to 3′, a segment of the inserted transgenic DNA (83nucleotides), and a segment of the 3′ genomic DNA adjacent to theinserted transgenic DNA (255 nucleotides) of event MON 87411.

SEQ ID NO:11 is a nucleotide sequence of a cDNA sequence from Diabroticavirgifera virgifera (Western Corn Rootworm) encoding an ESCRT-IIIcomplex subunit that is orthologous to yeast Snf7.

SEQ ID NO:12 is a nucleotide sequence representing the antisense strandof a DNA expression cassette that includes a recombinant gene engineeredto express an inverted repeat RNA molecule. The inverted repeat DNAsegments correspond to positions 663 through 902 and to positions 1292through 1053. The inverted repeat DNA sequences correspond to thenucleotide sequence of SEQ ID NO:11 from nucleotide position 151-390.

SEQ ID NO:13 is a ribonucleotide sequence transcribed from the DNA asset forth in SEQ ID NO:12.

SEQ ID NO:14 is a nucleotide sequence representing the sense strand of aDNA expression cassette that includes a recombinant gene engineered toencode and express a corn rootworm toxic Cry3Bb protein.

SEQ ID NO:15 is an amino acid sequence translation of a polynucleotidecorresponding to positions 1522-3480 of SEQ ID NO:14, and representing acorn rootworm toxic Cry3Bb protein.

SEQ ID NO:16 is a nucleotide sequence representing the sense strand of aDNA expression cassette that includes a recombinant gene engineered toencode and express a 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS)protein.

SEQ ID NO:17 is an amino acid sequence translation of a polynucleotidecorresponding to positions 2186 through 3781 of SEQ ID NO:16, andrepresenting an EPSPS protein that exhibits insensitivity to theherbicide glyphosate.

SEQ ID NO:18 is a nucleotide sequence of a synthetic oligonucleotidereferred to as SQ27011, and is identical to the nucleotide sequencecorresponding to positions 462-490 of SEQ ID NO:1.

SEQ ID NO:19 is a nucleotide sequence of a synthetic oligonucleotidereferred to as PB3552, and is identical to the reverse complement of thenucleotide sequence corresponding to positions 502-515 of SEQ ID NO:1.PB3552 can be 5′ labeled with a 6-carboxyfluorescein moiety (6-FAM™) and3′ labeled with a quencher moiety for use in combination with a pair ofthermal amplification primers, e.g., SQ27011 and SQ9085, and capable ofuse in TAQMAN® DNA amplification method to detect the presence of eventMON 87411 DNA in a biological sample that contains corn event MON 87411DNA.

SEQ ID NO:20 is a nucleotide sequence of a synthetic oligonucleotidereferred to as SQ9085, and is identical to the reverse complement of thenucleotide sequence corresponding to positions 516-541 of SEQ ID NO:1.

SEQ ID NO:21 is a nucleotide sequence of event MON 87411, andcorresponds to positions 462-541 of SEQ ID NO:1. An amplicon exhibitingthis sequence can be produced with a pair of thermal amplificationprimers, e.g., SQ27011 and SQ9085.

SEQ ID NO:22 is a nucleotide sequence of a synthetic oligonucleotidereferred to as SQ27066, and is identical to the nucleotide sequencecorresponding to positions 11710-11728 of SEQ ID NO:1.

SEQ ID NO:23 is a nucleotide sequence of a synthetic oligonucleotidereferred to as PB11300, and is identical to the nucleotide sequencecorresponding to positions 11731-11755 of SEQ ID NO:1. PB11300 can be 5′labeled with a 6-carboxyfluorescein moiety (6-FAM™) and 3′ labeled witha quencher moiety. Labeled this way, PB11300 can be used in combinationwith a pair of PCR primers, e.g., SQ27066 and SQ26977, to detect eventMON 87411 in a TAQMAN® assay.

SEQ ID NO:24 is a nucleotide sequence of a synthetic oligonucleotidereferred to as SQ26977, and is identical to the reverse complement ofthe nucleotide sequence corresponding to positions 11756-11784 of SEQ IDNO:1.

SEQ ID NO:25 is a nucleotide sequence of event MON 87411, andcorresponds to positions 11710-11784 of SEQ ID NO:1. An ampliconexhibiting this sequence can be amplified with a pair of primers, e.g.SQ27066 and SQ26977, and is diagnostic of event MON 87411.

SEQ ID NO:26 is a nucleotide sequence representing the DNA construct#417.

SEQ ID NO:27 is a nucleotide sequence representing the DNA construct#416.

SEQ ID NO:28 is a nucleotide sequence representing the DNA construct#418.

SEQ ID NO:29 is a nucleotide sequence representing the DNA construct#419.

SEQ ID NO:30 is a nucleotide sequence representing the DNA construct#402.

SEQ ID NO:31 is a nucleotide sequence representing the DNA construct#403.

SEQ ID NO:32 is a nucleotide sequence representing the DNA construct#404.

SEQ ID NO:33 is a nucleotide sequence representing the DNA construct#423.

SEQ ID NO:34 is a nucleotide sequence representing the DNA construct#405.

SEQ ID NO:35 is a nucleotide sequence representing the DNA construct#406.

SEQ ID NO:36 is a nucleotide sequence representing the DNA construct#890.

SEQ ID NO:37 is a nucleotide sequence of the LH244 corn plantrepresenting the wild-type allele of event MON 87411. An ampliconexhibiting this nucleotide sequence can be produced with a pair of PCRprimers, e.g., SQ27011 and SQ26977, and is diagnostic of the wild-typeallele of event MON 87411.

SEQ ID NO:38 is a nucleotide sequence of a synthetic oligonucleotidereferred to as SQ20221.

SEQ ID NO:39 is a nucleotide sequence of a synthetic oligonucleotidereferred to as PB10065. PB10065 can be 5′ labeled with VIC™ and 3′labeled with a quencher moiety. Labeled this way, PB10065 can be used incombination with a pair of PCR primers, e.g., SQ10065 and SQ20222, todetect the presence of a segment of an endogenous gene of corn in aTAQMAN® assay.

SEQ ID NO:40 is a nucleotide sequence of a synthetic oligonucleotidereferred to as SQ20222.

SEQ ID NOs:41-52 are nucleotide sequences of regions of SEQ ID NO:1,where each SEQ ID NO: encompasses a junction formed by interveningsequence and the expression cassette elements as detailed in the briefdescription for FIG. 3 .

DETAILED DESCRIPTION

The inventors have identified a transgenic corn event MON 87411exhibiting superior properties and performance compared to existingtransgenic corn plants. The corn event MON 87411 contains three operablylinked expression cassettes which collectively confer the traits of cornrootworm resistance and glyphosate herbicide tolerance to corn cells,corn tissues, corn seed and corn plants containing the transgenic eventMON 87411. The corn event MON 87411 provides two modes of action againstcorn rootworm pest species (including Diabrotica spp., especially whenthe pest is Diabrotica virgifera virgifera (Western Corn Rootworm, WCR),Diabrotica barberi (Northern Corn Rootworm, NCR), Diabrotica virgiferazeae (Mexican Corn Rootworm, MCR), Diabrotica balteata (Brazilian CornRootworm (BZR) or Brazilian Corn Rootworm complex (BCR) consisting ofDiabrotica viridula and Diabrotica speciosa), or Diabroticaundecimpunctata howardii (Southern Corn Rootworm, SCR)). Othertransgenic corn events have been referenced in the art that providevarious embodiments conferred singly, such as MON863 (conferring thetrait of resistance to corn rootworms by expression of a Cry3Bbinsecticidal toxin protein), or transgenic corn events providing two ormore traits such as in corn event MON88017 (conferring the trait ofresistance to corn rootworms by expression of a Cry3Bb insecticidaltoxin protein and the trait of resistance to glyphosate herbicide byexpression of a glyphosate insensitive EPSPS) and corn event DAS 59122-7(conferring the trait of resistance to corn rootworms by expression of abinary Bacillus thuringiensis toxin PS149B1, also known as Cry34/Cry35,and the trait of tolerance to the herbicide glufosinate). Other artdiscloses the combination by breeding of the traits conferred by thecorn events MON88017 or DAS 59122-7 with a transgenic corn eventconferring the trait of corn rootworm resistance resulting from theexpression of a dsRNA targeting for suppression a corn rootworm geneessential for the rootworms' survival (U.S. Pat. No. 7,943,819).Inherent in such combinations are the problems associated with the needfor breeding these multiple traits located in multiple different lociand on multiple chromosomes within the corn genome together into asingle corn plant and maintaining those traits as hybrids in dozens ifnot hundreds of different corn germplasm varieties. The solution forsuch problems would be to include combinations of these traits togetherin a single locus. The inventors herein provide one such solution to theproblem in the form of the corn event MON 87411, which combines threecovalently linked expression cassettes together in a single locus withinthe corn genome, these expression cassettes conferring the traits ofcorn rootworm resistance and glyphosate herbicide tolerance to the corncells, corn tissues, corn seed and corn plants containing the transgenicevent MON87411. Use of corn event MON 87411 provides major benefits tocorn growers: a) protection from economic losses due to the cornrootworm larvae by providing two different corn rootworm resistancemodes of action, and b) the ability to apply glyphosate containingagricultural herbicides to the corn crop for broad-spectrum weedcontrol. Additionally, the transgenes encoding the corn rootworm andglyphosate tolerant traits are linked on the same DNA segment and occurat a single locus in the genome of MON 87411, providing for enhancedbreeding efficiency and enables the use of molecular markers to trackthe transgene insert in the breeding populations and progeny thereof.

The corn event MON 87411 was produced by an Agrobacterium mediatedtransformation process of an inbred corn line with the plasmid constructpMON120417. This plasmid construct contains the linked plant expressioncassettes with the regulatory genetic elements necessary for expressionin corn plant cells of a CP4 EPSPS protein, as well as a Cry3Bb proteinand a dsRNA targeting for suppression an essential gene in the cells ofcorn rootworms when corn cells containing corn event MON 87411 areprovided in the diet of such corn rootworms. Corn cells were regeneratedinto intact corn plants and individual plants were selected from thepopulation of plants that showed integrity of the plant expressioncassettes and resistance to glyphosate and corn rootworm larvae feedingdamage. A corn plant that contains in its genome the linked plantexpression cassettes present in corn event MON 87411 is an aspect of thepresent invention.

The plasmid DNA inserted into the genome of corn event MON 87411 wascharacterized by detailed molecular analyses. These analyses included:the insert number (number of integration sites within the corn genome),the copy number (the number of copies of the T-DNA within one locus),and the integrity of the transgenic inserted DNA. The plasmid constructcontaining the three linked expression cassettes inserted into the corngenome giving rise to the event MON 87411 contains multiple segments(junction sequences between elements used to build or construct theseveral expression cassettes) that are not known to appear naturally inthe corn genome nor in other vectors or transgenic events of corn orotherwise (for example, sequences as set forth in SEQ ID NO:1, SEQ IDNO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7,SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10; SEQ ID NO:12, SEQ ID NO:14, SEQID NO:16, SEQ ID NO:21, SEQ ID NO:25, SEQ ID NO: 41, SEQ ID NO: 42, SEQID NO: 43, SEQ ID NO: 44, SEQ ID NO: 45, SEQ ID NO: 49, SEQ ID NO: 50,SEQ ID NO: 51, and SEQ ID NO: 52). In addition, the transformation eventthat gave rise to the inserted transgenic DNA in the event MON 87411 ischaracterized herein as an insertion into a single locus in the corngenome, resulting in two new loci or junction sequences between theinserted DNA and the corn genome DNA (additional junction sequences)that are of sufficient length to be unique only to a corn genomecomprising event MON 87411. These junction sequences are useful fordetecting the presence of the event MON 87411 DNA in corn cells, tissue,seed and plants or plant products (commodity products). DNA molecularprobes and primer pairs are described herein that have been developedfor use in identifying the presence of these various junction segmentsin biological samples containing or suspected of containing corn cells,seed, plant parts or plant tissue that contain the event MON 87411 DNA.The data show that event MON 87411 contains a single T-DNA insertionwith one copy of the inserted transgenic DNA. No additional elementsfrom the transformation vector pMON120714 other than portions of theAgrobacterium tumefaciens left and right border regions used fortransgenic DNA transfer from the plant transformation plasmid to thecorn genome have been identified in event MON 87411. Finally, thermalamplification producing specific amplicons diagnostic for the presenceof such event MON 87411 DNA in a sample, and DNA sequence analyses wereperformed to determine the arbitrarily assigned 5′ and 3′insert-to-plant genome junctions, confirm the organization of theelements within the insert, and determine the complete DNA sequence ofthe inserted transgene DNA in corn plant event MON 87411 (SEQ ID NO:1).

Dozens of transgenic events were produced using the construct used toproduce the transgenic event MON 87411, and different constructs wereproduced and used to produce many dozens of other transgenic corn eventswhich were compared to the MON 87411 and similar events. These eventswere all tested for efficacy for controlling corn rootworms in dietbioassays in which the transgenic corn plant event tissues were providedin the diet of corn rootworm larvae. It was determined that theorientation of expression of the two different expression cassettesresponsible for conferring the corn rootworm resistance traits to thevarious events was critical to the efficacy of the events in providingcorn rootworm control when the corn event cells expressing theseresistance traits were provided in the diet of the corn rootworm larvae.Two different promoters, CAMV e35S and Zm.PIIG, were observed to providesurprising and superior efficacy of corn events containing expressioncassettes expressing the dsRNA corn rootworm protectant from the e35Spromoter and the Cry3Bb corn rootworm toxic protein from an a Zm.PIIGpromoter that was adjacent to and divergent from the e35S promoter. Whenthese promoters were in this particular orientation significantlyimproved ratios of transgenic events exhibiting efficacy were obtained.

Unless otherwise noted herein, terms are to be understood according toconventional usage by those of ordinary skill in the relevant art.Definitions of common terms in molecular biology may also be found inRieger et al., Glossary of Genetics: Classical and Molecular, 5thedition, Springer-Verlag: New York, 1991; and Lewin, Genes V, OxfordUniversity Press: New York, 1994. As used herein, the term “corn” meansZea mays and includes all plant varieties that can be bred with cornplants comprising MON 87411. As used herein, the term “comprising” means“including but not limited to”.

The present invention provides for transgenic plants which have beentransformed with a DNA construct that contains at least three expressioncassettes; a first expression cassette expressing a corn rootworm toxicamount of a dsRNA designed to suppress a corn rootworm essential geneorthologous to a yeast snf7 gene, a second expression cassette expressescorn rootworm toxic amounts of Cry3Bb delta-endotoxin, and a thirdexpression cassette that expresses a glyphosate tolerance enzyme CP4EPSPS that is insensitive to glyphosate inhibition. Corn plantstransformed according to the methods and with the DNA constructdisclosed herein are resistant to CRW and tolerant to applications ofglyphosate herbicide. The linked agronomic traits provide ease inmaintaining these traits together in a breeding population, and exhibitgreater corn rootworm efficacy than plants containing only a single cornrootworm inhibition gene or that contain the same corn rootworminhibition genes (Cry3Bb and dsRNA) that are combined as a breedingstack.

A transgenic “plant” is produced by transformation of a plant cell withheterologous DNA, i.e., a polynucleic acid construct that includes atransgene of interest; regeneration of a population of plants resultingfrom the insertion of the transgene into the genome of the plant cell,and selection of a particular plant characterized by insertion into aparticular genome location. The term “event” refers to the originaltransformant plant and progeny of the transformant that include theheterologous DNA. The term “event” also includes progeny produced by asexual outcross between the event and another plant wherein the progenyincludes the heterologous DNA. Even after repeated back-crossing to arecurrent parent, the inserted DNA and flanking genomic DNA from thetransformed parent event is present in the progeny of the cross at thesame chromosomal location. The term “event” also refers to DNA from theoriginal transformant comprising the inserted DNA, and flanking genomicsequence immediately adjacent to the inserted DNA, that would beexpected to be transferred to a progeny that receives the inserted DNAincluding the transgene of interest as the result of a sexual cross ofone parental line that includes the inserted DNA (e.g., the originaltransformant and progeny resulting from selfing) and a parental linethat does not contain the inserted DNA. The present invention is relatedto the transgenic event, corn plant comprising MON 87411, progenythereof, and DNA compositions contained therein.

A “probe” is an isolated nucleic acid to which is attached aconventional detectable label or reporter molecule, e.g., a radioactiveisotope, ligand, chemiluminescent agent, or enzyme. Such a probe iscomplementary to a strand of a target nucleic acid, in the case of thepresent invention, to a strand of genomic DNA from MON 87411 whetherfrom a MON 87411 plant or from a sample that includes MON 87411 DNA.Probes according to the present invention include not onlydeoxyribonucleic or ribonucleic acids, but also polyamides and otherprobe materials that bind specifically to a target DNA sequence and canbe used to detect the presence of that target DNA sequence.

DNA primers are isolated polynucleic acids that are annealed to acomplementary target DNA strand by nucleic acid hybridization to form ahybrid between the primer and the target DNA strand, then extended alongthe target DNA strand by a polymerase, e.g., a DNA polymerase. A DNAprimer pair or a DNA primer set of the present invention refer to twoDNA primers useful for amplification of a target nucleic acid sequence,e.g., by the polymerase chain reaction (PCR) or other conventionalpolynucleic acid amplification methods.

DNA probes and DNA primers are generally 11 polynucleotides or more inlength, often 18 polynucleotides or more, 24 polynucleotides or more, or30 polynucleotides or more. Such probes and primers are selected to beof sufficient length to hybridize specifically to a target sequenceunder high stringency hybridization conditions. Preferably, probes andprimers according to the present invention have complete sequencesimilarity with the target sequence, although probes differing from thetarget sequence that retain the ability to hybridize to target sequencesmay be designed by conventional methods.

Primers and probes based on the flanking genomic DNA and insertsequences disclosed herein can be used to confirm (and, if necessary, tocorrect) the disclosed DNA sequences by conventional methods, e.g., byre-cloning and sequencing such DNA molecules.

The nucleic acid probes and primers of the present invention hybridizeunder stringent conditions to a target DNA molecule. Any conventionalnucleic acid hybridization or amplification method can be used toidentify the presence of DNA from a transgenic plant in a sample.Polynucleic acid molecules, also referred to as nucleic acid segments,or fragments thereof are capable of specifically hybridizing to othernucleic acid molecules under certain circumstances. As used herein, twopolynucleic acid molecules are said to be capable of specificallyhybridizing to one another if the two molecules are capable of formingan anti-parallel, double-stranded nucleic acid structure. A nucleic acidmolecule is said to be the “complement” of another nucleic acid moleculeif they exhibit complete complementarity. As used herein, molecules aresaid to exhibit “complete complementarity” when every nucleotide of oneof the molecules is complementary to a nucleotide of the other. Twomolecules are said to be “minimally complementary” if they can hybridizeto one another with sufficient stability to permit them to remainannealed to one another under at least conventional “low-stringency”conditions. Similarly, the molecules are said to be “complementary” ifthey can hybridize to one another with sufficient stability to permitthem to remain annealed to one another under conventional“high-stringency” conditions. Conventional stringency conditions aredescribed by Sambrook et al., 1989, and by Haymes et al., In: NucleicAcid Hybridization, A Practical Approach, IRL Press, Washington, D.C.(1985), Departures from complete complementarity are thereforepermissible, as long as such departures do not completely preclude thecapacity of the molecules to form a double-stranded structure. In orderfor a nucleic acid molecule to serve as a primer or probe it need onlybe sufficiently complementary in sequence to be able to form a stabledouble-stranded structure under the particular solvent and saltconcentrations employed.

As used herein, a substantially homologous sequence is a nucleic acidsequence that will specifically hybridize to the complement of thenucleic acid sequence to which it is being compared under highstringency conditions. Appropriate stringency conditions that promoteDNA hybridization, for example, 6.0× sodium chloride/sodium citrate(SSC) at about 45° C., followed by a wash of 2.0×SSC at 50° C., areknown to those skilled in the art or can be found in Current Protocolsin Molecular Biology, John Wiley & Sons, N.Y. (1989), 6.3.1-6.3.6. Forexample, the salt concentration in the wash step can be selected from alow stringency of about 2.0×SSC at 50° C. to a high stringency of about0.2×SSC at 50° C. In addition, the temperature in the wash step can beincreased from low stringency conditions at room temperature, about 22°C., to high stringency conditions at about 65° C. Both temperature andsalt may be varied, or either the temperature or the salt concentrationmay be held constant while the other variable is changed. In a preferredembodiment, a polynucleic acid of the present invention willspecifically hybridize to one or more of the nucleic acid molecules setforth in SEQ ID NOs: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 12, 14, 16, 21, 25,41, 42, 43, 44, 45, 49, 50, 51, or 52 or complements thereof orfragments of either under moderately stringent conditions, for exampleat about 2.0×SSC and about 65° C. In a particularly preferredembodiment, a nucleic acid of the present invention will specificallyhybridize to one or more of the nucleic acid molecules set forth in SEQID NOs: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 12, 14, 16, 21, 25, 41, 42, 43,44, 45, 49, 50, 51, or 52 or complements or fragments of either underhigh stringency conditions. In one aspect of the present invention, apreferred marker nucleic acid molecule of the present invention has thenucleic acid sequence set forth in SEQ ID NO:1, or SEQ ID NO:2, or SEQID NO:3, or SEQ ID NO:4, or SEQ ID NO:5, or SEQ ID NO:6, or SEQ ID NO:7,or SEQ ID NO:8, or SEQ ID NO:9, or SEQ ID NO:10; or SEQ ID NO:12, or SEQID NO:14, OR SEQ ID NO:16, or SEQ ID NO:21, or SEQ ID NO:25, or SEQ IDNO: 41, or SEQ ID NO: 42, or SEQ ID NO: 43, or SEQ ID NO: 44, or SEQ IDNO: 45, or SEQ ID NO: 49, or SEQ ID NO: 50, or SEQ ID NO: 51, or SEQ IDNO: 52 or complements thereof or fragments of either. The hybridizationof the probe to the target DNA molecule can be detected by any number ofmethods known to those skilled in the art, these can include, but arenot limited to, fluorescent tags, radioactive tags, antibody based tags,and chemiluminescent tags.

Regarding the amplification of a target nucleic acid sequence (e.g., byPCR) using a particular amplification primer pair, “stringentconditions” are conditions that permit the primer pair to hybridize onlyto the target nucleic acid sequence to which a primer having thecorresponding wild-type sequence (or its complement) would bind andpreferably to produce a unique amplification product, the amplicon, in aDNA thermal amplification reaction.

The term “specific for (a target sequence)” indicates that a probe orprimer hybridizes under stringent hybridization conditions only to thetarget sequence in a sample comprising the target sequence.

As used herein, “amplified DNA” or “amplicon” refers to the product ofpolynucleic acid amplification method directed to a target polynucleicacid molecule that is part of a polynucleic acid template. For example,to determine whether a corn plant resulting from a sexual cross containstransgenic plant genomic DNA from a corn plant comprising MON 87411 ofthe present invention, DNA that is extracted from a corn plant tissuesample may be subjected to a polynucleic acid amplification method usinga primer pair that includes a primer derived from a DNA sequence in thegenome of a MON 87411 comprising plant adjacent to the insertion site ofthe inserted heterologous DNA (transgene DNA), and a second primerderived from the inserted heterologous DNA to produce an amplicon thatis diagnostic for the presence of the MON 87411 plant DNA. Thediagnostic amplicon is of a length and has a DNA sequence that is alsodiagnostic for the plant genomic DNA. The amplicon may range in lengthfrom the combined length of the primer pair plus one nucleotide basepair, preferably plus about fifty nucleotide base pairs, more preferablyplus about two hundred-fifty nucleotide base pairs, and even morepreferably plus about four hundred-fifty nucleotide base pairs or more.Alternatively, a primer pair can be derived from genomic sequence onboth sides of the inserted heterologous DNA so as to produce an ampliconthat includes the entire insert polynucleotide sequence (e.g., a forwardprimer isolated from the genomic portion of SEQ ID NO:1 and a reverseprimer isolated from the genomic portion of SEQ ID NO:1 that amplifies aDNA molecule comprising the a junction sequence identified herein in theevent MON 87411 genome). A member of a primer pair derived from theplant genomic sequence adjacent to the inserted transgenic DNA islocated a distance from the inserted DNA sequence, this distance canrange from one nucleotide base pair up to about twenty thousandnucleotide base pairs. The use of the term “amplicon” specificallyexcludes primer dimers that may be formed in the DNA thermalamplification reaction.

Polynucleic acid amplification can be accomplished by any of the variouspolynucleic acid amplification methods known in the art, including thepolymerase chain reaction (PCR). Amplification methods are known in theart and are described, inter alia, in U.S. Pat. Nos. 4,683,195 and4,683,202 and in PCR Protocols: A Guide to Methods and Applications, ed.Innis et al., Academic Press, San Diego, 1990. PCR amplification methodshave been developed to amplify up to 22 kb (kilobase) of genomic DNA andup to 42 kb of bacteriophage DNA (Cheng et al., Proc. Natl. Acad. Sci.USA 91:5695-5699, 1994). These methods as well as other methods known inthe art of DNA amplification may be used in the practice of the presentinvention. The sequence of the heterologous DNA insert or flankinggenomic DNA sequence from event MON 87411 can be verified (and correctedif necessary) by amplifying such DNA molecules from event MON 87411comprising seed or plants grown from the seed deposited with the ATCChaving accession no. PTA-12669, using primers derived from the sequencesprovided herein, followed by standard DNA sequencing of the PCR ampliconor cloned DNA fragments thereof.

DNA detection kits that are based on DNA amplification methods containDNA primer molecules that hybridize specifically to a target DNA andamplify a diagnostic amplicon under the appropriate reaction conditions.The kit may provide an agarose gel based detection method or any numberof methods of detecting the diagnostic amplicon that are known in theart. A kit that contains DNA primers that are homologous orcomplementary to any portion of the corn genomic region as set forth inSEQ ID NO:1 and to any portion of the inserted transgenic DNA as setforth in SEQ ID NO:1 is an object of the invention. DNA molecules usefulas DNA primers can be selected from the disclosed transgene/genomic DNAsequence of MON 87411 (SEQ ID NO:1) by those skilled in the art of DNAamplification.

The diagnostic amplicon produced by these methods may be detected by aplurality of techniques. One such method is Genetic Bit Analysis(Nikiforov, et al. Nucleic Acid Res. 22:4167-4175, 1994) where a DNAoligonucleotide is designed that overlaps both the adjacent flankinggenomic DNA sequence and the inserted DNA sequence. The oligonucleotideis immobilized in wells of a microtiter plate. Following PCR of theregion of interest (using one primer in the inserted sequence and one inthe adjacent flanking genomic sequence), a single-stranded PCR productcan be hybridized to the immobilized oligonucleotide and serve as atemplate for a single base extension reaction using a DNA polymerase andlabeled dideoxynucleotide triphosphates (ddNTPs) specific for theexpected next base. Readout may be fluorescent or ELISA-based. A signalindicates presence of the transgene/genomic sequence due to successfulamplification, hybridization, and single base extension.

Another method is the Pyrosequencing technique as described by Winge(Innov. Pharma. Tech. 00:18-24, 2000). In this method an oligonucleotideis designed that overlaps the adjacent genomic DNA and insert DNAjunction. The oligonucleotide is hybridized to single-stranded PCRproduct from the region of interest (one primer in the inserted sequenceand one in the flanking genomic sequence) and incubated in the presenceof a DNA polymerase, ATP, sulfurylase, luciferase, apyrase, adenosine 5′phosphosulfate and luciferin. DNTPs are added individually and theincorporation results in a light signal that is measured. A light signalindicates the presence of the transgene/genomic sequence due tosuccessful amplification, hybridization, and single or multi-baseextension.

Fluorescence Polarization as described by Chen, et al., (Genome Res.9:492-498, 1999) is a method that can be used to detect the amplicon ofthe present invention. Using this method an oligonucleotide is designedthat overlaps the genomic flanking and inserted DNA junction. Theoligonucleotide is hybridized to single-stranded PCR product from theregion of interest (one primer in the inserted DNA and one in theflanking genomic DNA sequence) and incubated in the presence of a DNApolymerase and a fluorescent-labeled ddNTP. Single base extensionresults in incorporation of the ddNTP. Incorporation can be measured asa change in polarization using a fluorometer. A change in polarizationindicates the presence of the transgene/genomic sequence due tosuccessful amplification, hybridization, and single base extension.

Taqman® (PE Applied Biosystems, Foster City, CA) is described as amethod of detecting and quantifying the presence of a DNA sequence andis fully understood in the instructions provided by the manufacturer.Briefly, a FRET oligonucleotide probe is designed that overlaps thegenomic flanking and insert DNA junction. The FRET probe and PCR primers(one primer in the insert DNA sequence and one in the flanking genomicsequence) are cycled in the presence of a thermostable polymerase anddNTPs. Hybridization of the FRET probe results in cleavage and releaseof the fluorescent moiety away from the quenching moiety on the FRETprobe. A fluorescent signal indicates the presence of thetransgene/genomic sequence due to successful amplification andhybridization.

Molecular Beacons have been described for use in sequence detection asdescribed in Tyangi, et al. (Nature Biotech. 14:303-308, 1996) Briefly,a FRET oligonucleotide probe is designed that overlaps the flankinggenomic and insert DNA junction. The unique structure of the FRET proberesults in it containing secondary structure that keeps the fluorescentand quenching moieties in close proximity. The FRET probe and PCRprimers (one primer in the insert DNA sequence and one in the flankinggenomic sequence) are cycled in the presence of a thermostablepolymerase and dNTPs. Following successful PCR amplification,hybridization of the FRET probe to the target sequence results in theremoval of the probe secondary structure and spatial separation of thefluorescent and quenching moieties. A fluorescent signal results. Afluorescent signal indicates the presence of the flanking/transgeneinsert sequence due to successful amplification and hybridization.

DNA detection kits can be developed using the compositions disclosedherein and the methods well known in the art of DNA detection. The kitsare useful for identification of corn event MON 87411 DNA in a sampleand can be applied to methods for breeding corn plants containing MON87411 DNA. A kit contains DNA molecules that are useful as primers orprobes and that are homologous or complementary to at least theapplicable portions of SEQ ID NO:1 as described herein. The DNAmolecules can be used in DNA amplification methods (PCR) or as probes inpolynucleic acid hybridization methods, i.e., Southern analysis,northern analysis.

Junction sequences may be represented by a sequence from the groupconsisting of SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:5, SEQ ID NO:6, SEQ IDNO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10; SEQ ID NO:21, SEQ IDNO:25, SEQ ID NO:41, SEQ ID NO:42, SEQ ID NO:43, SEQ ID NO:44, SEQ IDNO:45, SEQ ID NO:49, SEQ ID NO:50, SEQ ID NO:51, and SEQ ID NO:52. Forexample, the junction sequences may be arbitrarily represented by thenucleotide sequences provided as SEQ ID NO:5 and SEQ ID NO:8.Alternatively, the junction sequences may be arbitrarily represented bythe nucleotide sequences provided as SEQ ID NO:6 and SEQ ID NO:9.Alternatively, the junction sequences may be arbitrarily represented bythe nucleotide sequences provided as SEQ ID NO:7 and SEQ ID NO:10. Thesenucleotides are connected by phosphodiester linkage and in corn eventMON 87411 are present as part of the recombinant plant cell genome. Theidentification of one or more of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3,SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ IDNO:9, SEQ ID NO:10, SEQ ID NO:2, SEQ ID NO:25, SEQ ID NO:41, SEQ IDNO:42, SEQ ID NO:43, SEQ ID NO:44, SEQ ID NO:45, SEQ ID NO:49, SEQ IDNO:50, SEQ ID NO:51, or SEQ ID NO:52 in a sample derived from a cornplant, seed, or plant part is determinative that the DNA was obtainedfrom corn event MON 87411 and is diagnostic for the presence in a samplecontaining DNA from corn event MON 87411. The invention thus provides aDNA molecule that contains at least one of the nucleotide sequencesprovided as SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ IDNO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10,SEQ ID NO:21, SEQ ID NO:25, SEQ ID NO:41, SEQ ID NO:42, SEQ ID NO:43,SEQ ID NO:44, SEQ ID NO:45, SEQ ID NO:49, SEQ ID NO:50, SEQ ID NO:51, orSEQ ID NO:52. Any segment of DNA derived from transgenic corn event MON87411 that is sufficient to include at least one of the sequencesprovided as SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ IDNO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10,SEQ ID NO:21, SEQ ID NO:25, SEQ ID NO:41, SEQ ID NO:42, SEQ ID NO:43,SEQ ID NO:44, SEQ ID NO:45, SEQ ID NO:49, SEQ ID NO:50, SEQ ID NO:51, orSEQ ID NO:52 is within the scope of the invention. In addition, anypolynucleotide comprising a sequence complementary to any of thesequences described within this paragraph is within the scope of theinvention.

The invention provides exemplary DNA molecules that can be used eitheras primers or probes for detecting the presence of DNA derived from acorn plant comprising event MON 87411 DNA in a sample. Such primers orprobes are specific for a target nucleic acid sequence and as such areuseful for the identification of corn event MON 87411 nucleic acidsequence by the methods of the invention described herein.

A “primer” is typically a highly purified, isolated polynucleotide thatis designed for use in specific annealing or hybridization methods thatinvolve thermal amplification. A pair of primers may be used withtemplate DNA, such as a sample of corn genomic DNA, in a thermalamplification, such as polymerase chain reaction (PCR), to produce anamplicon, where the amplicon produced from such reaction would have aDNA sequence corresponding to sequence of the template DNA locatedbetween the two sites where the primers hybridized to the template. Asused herein, an “amplicon” is a piece or fragment of DNA that has beensynthesized using amplification techniques. An amplicon of the inventioncomprises at least one of the sequences provided as SEQ ID NO:21 or SEQID NO:25. A primer is typically designed to hybridize to a complementarytarget DNA strand to form a hybrid between the primer and the target DNAstrand, and the presence of the primer is a point of recognition by apolymerase to begin extension of the primer (i.e., polymerization ofadditional nucleotides into a lengthening nucleotide molecule) using asa template the target DNA strand. Primer pairs, as used in theinvention, are intended to refer to the use of two primers bindingopposite strands of a double stranded nucleotide segment for the purposeof amplifying linearly the polynucleotide segment between the positionstargeted for binding by the individual members of the primer pair,typically in a thermal amplification reaction or other conventionalnucleic-acid amplification methods. A primer pair useful for thisapplication should comprise a first DNA molecule and a second DNAmolecule that is different from the first DNA molecule, and wherein bothare each of sufficient length of contiguous nucleotides of a DNAsequence to function as DNA primers that, when used together in athermal amplification reaction with template DNA derived from corn eventMON 87411, to produce an amplicon diagnostic for corn event MON 87411DNA in a sample. Exemplary DNA molecules useful as primers are providedas SEQ ID NO:18, SEQ ID NO:20, SEQ ID NO:22, or SEQ ID NO:24.

A “probe” is an isolated nucleic acid that is complementary to a strandof a target nucleic acid. Probes include not only deoxyribonucleic orribonucleic acids but also polyamides and other probe materials thatbind specifically to a target DNA sequence and the detection of suchbinding can be useful in diagnosing, discriminating, determining,detecting, or confirming the presence of that target DNA sequence in aparticular sample. A probe may be attached to a conventional detectablelabel or reporter molecule, e.g., a radioactive isotope, ligand,chemiluminescent agent, or enzyme. Exemplary DNA molecules useful asprobes are provided as SEQ ID NO:19 and SEQ ID NO:23.

Probes and primers may have complete sequence identity with the targetsequence, although primers and probes differing from the target sequencethat retain the ability to hybridize preferentially to target sequencesmay be designed by conventional methods. In order for a nucleic acidmolecule to serve as a primer or probe it need only be sufficientlycomplementary in sequence to be able to form a stable double-strandedstructure under the particular solvent and salt concentrations employed.Any conventional nucleic acid hybridization or amplification method canbe used to identify the presence of transgenic DNA from corn event MON87411 in a sample. Probes and primers are generally at least about 11nucleotides, at least about 18 nucleotides, at least about 24nucleotides, or at least about 30 nucleotides or more in length. Suchprobes and primers hybridize specifically to a target DNA sequence understringent hybridization conditions. Conventional stringency conditionsare described by Sambrook et al., 1989, and by Haymes et al., In:Nucleic Acid Hybridization, A Practical Approach, IRL Press, Washington,D.C. (1985).

Any number of methods well known to those skilled in the art can be usedto isolate and manipulate a DNA molecule, or fragment thereof, disclosedin the invention, including thermal amplification methods. DNAmolecules, or fragments thereof, can also be obtained by othertechniques such as by directly synthesizing the fragment by chemicalmeans, as is commonly practiced by using an automated oligonucleotidesynthesizer.

The DNA molecules and corresponding nucleotide sequences provided hereinare therefore useful for, among other things, identifying corn event MON87411, selecting plant varieties or hybrids comprising corn event MON87411, detecting the presence of DNA derived from the transgenic cornevent MON 87411 in a sample, and monitoring samples for the presenceand/or absence of corn event MON 87411 or plant parts derived from cornplants comprising event MON 87411.

The invention provides corn plants, progeny, seeds, plant cells, plantparts (such as pollen, ovule, ear or silk tissue, tassel tissue, roottissue, stem tissue, and leaf tissue), and commodity products. Theseplants, progeny, seeds, plant cells, plant parts, and commodity productscontain a detectable amount of a polynucleotide of the invention, i.e.,such as a polynucleotide having at least one of the sequences providedas SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, SEQ IDNO:12, SEQ ID NO:14, SEQ ID NO:16, SEQ ID NO:21, SEQ ID NO:25, SEQ IDNO:41, SEQ ID NO:42, SEQ ID NO:43, SEQ ID NO:44, SEQ ID NO:45, SEQ IDNO:49, SEQ ID NO:50, SEQ ID NO:51, or SEQ ID NO:52. Plants, progeny,seeds, plant cells, and plant parts of the invention may also containone or more additional transgenes. Such additional transgene may be anynucleotide sequence encoding a protein or RNA molecule conferring adesirable trait including but not limited to increased insectresistance, increased water use efficiency, increased yield performance,increased drought resistance, increased seed quality, improvednutritional quality, and/or increased herbicide tolerance, in which thedesirable trait is measured with respect to a corn plant lacking suchadditional transgene.

The invention provides corn plants, progeny, seeds, plant cells, andplant part such as pollen, ovule, ear or silk tissue, tassel tissue,root or stem tissue, and leaves derived from a transgenic corn plantcomprising event MON 87411. A representative sample of corn seedcomprising event MON 87411 has been deposited according to the BudapestTreaty with the American Type Culture Collection (ATCC). The ATCCdepository has assigned the Patent Deposit Designation PTA-12669 to theevent MON 87411 comprising seed.

The invention provides a microorganism comprising a DNA molecule havingat least one sequence selected from the group consisting of SEQ ID NO:1,SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ IDNO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, SEQ ID NO:12, SEQ IDNO:14, SEQ ID NO:16, SEQ ID NO:21, SEQ ID NO:25, SEQ ID NO:41, SEQ IDNO:42, SEQ ID NO:43, SEQ ID NO:44, SEQ ID NO:45, SEQ ID NO:49, SEQ IDNO:50, SEQ ID NO:51, and SEQ ID NO:52 present in its genome. An exampleof such a microorganism is a transgenic plant cell. Microorganisms, suchas a plant cell of the invention, are useful in many industrialapplications, including but not limited to: (i) use as research tool forscientific inquiry or industrial research; (ii) use in culture forproducing endogenous or recombinant carbohydrate, lipid, nucleic acid,or protein products or small molecules that may be used for subsequentscientific research or as industrial products; and (iii) use with modernplant tissue culture techniques to produce transgenic plants or planttissue cultures that may then be used for agricultural research orproduction. The production and use of microorganisms such as transgenicplant cells utilizes modern microbiological techniques and humanintervention to produce a man-made, unique microorganism. In thisprocess, recombinant DNA is inserted into a plant cell's genome tocreate a transgenic plant cell that is separate and unique fromnaturally occurring plant cells. This transgenic plant cell can then becultured much like bacteria and yeast cells using modern microbiologytechniques and may exist in an undifferentiated, unicellular state. Thetransgenic plant cell's new or altered genetic composition and phenotypeis a technical effect created by the integration of the heterologous DNAinto the genome of the cell. Microorganisms of the invention, such astransgenic plant cells, include (i) methods of producing transgeniccells by integrating recombinant DNA into the genome of the cell andthen using this cell to derive additional cells possessing the sameheterologous DNA; (ii) methods of culturing cells that containrecombinant DNA using modern microbiology techniques; (iii) methods ofproducing and purifying endogenous or recombinant carbohydrate, lipid,nucleic acid, or protein products from cultured cells; and (iv) methodsof using modern plant tissue culture techniques with transgenic plantcells to produce transgenic plants or transgenic plant tissue cultures.

Plants of the invention may pass along the event DNA, including thetransgene, to progeny. As used herein, “progeny” includes any plant,seed, plant cell, and/or regenerable plant part comprising the event DNAderived from an ancestor plant and/or comprising a DNA molecule havingat least one sequence selected from the group consisting of SEQ ID NO:1,SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ IDNO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, SEQ ID NO:12, SEQ IDNO:14, SEQ ID NO:16, SEQ ID NO:21, SEQ ID NO:25; SEQ ID NO: 41, SEQ IDNO: 42, SEQ ID NO: 43, SEQ ID NO: 44, SEQ ID NO: 45, SEQ ID NO: 49, SEQID NO: 50, SEQ ID NO: 51, and SEQ ID NO:52. Plants, progeny, and seedsmay be homozygous or heterozygous for the transgene. Progeny may begrown from seeds produced by a corn event MON 87411 containing plantand/or from seeds produced by a plant fertilized with pollen from a cornevent MON 87411 containing plant.

Progeny plants may be self-pollinated (also known as “selfing”) togenerate a true breeding line of plants, i.e., plants homozygous for thetransgene. Selfing of appropriate progeny can produce plants that arehomozygous for both added, exogenous genes.

Alternatively, progeny plants may be outcrossed, e.g., bred with anotherunrelated plant, to produce a varietal or a hybrid seed or plant. Theother unrelated plant may be transgenic or nontransgenic. A varietal orhybrid seed or plant of the invention may thus be derived by crossing afirst parent that lacks the specific and unique DNA of the corn eventMON 87411 with a second parent comprising corn event MON 87411,resulting in a hybrid comprising the specific and unique DNA of the cornevent MON 87411. Each parent can be a hybrid or an inbred/varietal, solong as the cross or breeding results in a plant or seed of theinvention, i.e., a seed having at least one allele containing the DNA ofcorn event MON 87411 and/or a DNA molecule having at least one sequenceselected from the group consisting of SEQ ID NO:1, SEQ ID NO:2, SEQ IDNO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8,SEQ ID NO:9, SEQ ID NO:10, SEQ ID NO:12, SEQ ID NO:14, SEQ ID NO:16, SEQID NO:21, SEQ ID NO:25; SEQ ID NO: 41, SEQ ID NO: 42, SEQ ID NO: 43, SEQID NO: 44, SEQ ID NO: 45, SEQ ID NO: 49, SEQ ID NO: 50, SEQ ID NO: 51,and SEQ ID NO: 52. Two different transgenic plants may thus be crossedto produce hybrid offspring that contain two independently segregating,added, exogenous genes. For example, the event MON 87411 corn containingresistance to corn rootworm infestations and glyphosate tolerance can becrossed with different transgenic corn plants to produce a hybrid orinbred plant having the characteristics of both transgenic parents. Oneexample of this would be a cross of event MON 87411 containingresistance to corn rootworm infestations and glyphosate tolerance with acorn plant having one or more additional traits such as herbicidetolerance and/or insect control, resulting in a progeny plant or seedthat is resistant to corn rootworm infestations and tolerant toglyphosate and has at least one or more additional traits. Back-crossingto a parental plant and out-crossing with a non-transgenic plant arealso contemplated, as is vegetative propagation. Descriptions of otherbreeding methods that are commonly used for different traits and cropscan be found in one of several references, e.g., Fehr, in BreedingMethods for Cultivar Development, Wilcox J. ed., American Society ofAgronomy, Madison WI (1987).

The invention provides a plant part that is derived from corn plantscomprising event MON 87411. As used herein, a “plant part” refers to anypart of a plant which is comprised of material derived from a corn plantcomprising event MON 87411. Plant parts include but are not limited topollen, ovule, ear or silk, tassel, root or stem tissue, fibers, andleaves. Plant parts may be viable, nonviable, regenerable, and/ornonregenerable.

The invention provides a commodity product that is derived from cornplants comprising event MON 87411 and that contains a detectable amountof a nucleic acid specific for event MON 87411. As used herein, a“commodity product” refers to any composition or product which containsmaterial derived from a corn plant, whole or processed corn seed, one ormore plant cells and/or plant parts containing the corn event MON 87411DNA. Commodity products may be sold to consumers and may be viable ornonviable. Nonviable commodity products include but are not limited tononviable corn seeds; processed corn seeds, corn seed parts, and cornplant parts; corn seeds and corn plant parts processed for feed or food,oil, meal, flour, flakes, bran, biomasses, and fuel products. Viablecommodity products include but are not limited to corn seeds, cornplants, and corn plant cells. The corn plants comprising event MON 87411can thus be used to manufacture any commodity product typically acquiredfrom corn. Any such commodity product that is derived from corn plantscontaining corn event MON 87411 DNA that contains at least a detectableamount of one or more specific and unique DNA molecules, the presence ofwhich are determinative of corn event MON 87411, and specifically maycontain a detectable amount of a polynucleotide comprising a DNAmolecule having at least one sequence selected from the group consistingof SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, SEQ IDNO:12, SEQ ID NO:14, SEQ ID NO:16, SEQ ID NO:21, SEQ ID NO:25; SEQ IDNO: 41, SEQ ID NO: 42, SEQ ID NO: 43, SEQ ID NO: 44, SEQ ID NO: 45, SEQID NO: 49, SEQ ID NO: 50, SEQ ID NO: 51, and SEQ ID NO:52. Any standardmethod of detection for nucleotide molecules may be used, includingmethods of detection disclosed herein. A commodity product is within thescope of the invention if there is any detectable amount of a DNAmolecule having at least one diagnostic sequence selected from the groupconsisting of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ IDNO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10,SEQ ID NO:12, SEQ ID NO:14, SEQ ID NO:16, SEQ ID NO:21, SEQ ID NO:25;SEQ ID NO: 41, SEQ ID NO: 42, SEQ ID NO: 43, SEQ ID NO: 44, SEQ ID NO:45, SEQ ID NO: 49, SEQ ID NO: 50, SEQ ID NO: 51, and SEQ ID NO: 52 inthe commodity product.

The plants, progeny, seeds, plant cells, plant parts (such as pollen,ovule, ear or silk, tassel, root or stem tissue, and leaves), andcommodity products of the invention are therefore useful for, amongother things, growing plants for the purpose of producing seed and/orplant parts comprising corn event MON 87411 for agricultural purposes,producing progeny comprising corn event MON 87411 for plant breeding andresearch purposes, use with microbiological techniques for industrialand research applications, and sale to consumers.

The invention provides methods for controlling weeds and methods forproducing plants using glyphosate herbicide and corn event MON 87411. Amethod for controlling weeds in a field is provided and consists ofplanting corn event MON 87411 containing varietal or hybrid plants in afield and applying a herbicidally effective dose of glyphosate to thefield for the purpose of controlling weeds in the field without injuringthe MON 87411 containing plants. Such application of glyphosateherbicide may be pre-emergence, i.e., any time after MON 87411containing seed is planted and before MON 87411 containing plantsemerge, or post-emergence, i.e., any time after MON 87411 containingplants emerge. Another method for controlling weeds in a field is alsoprovided and consists of applying an effective dose of glyphosateherbicide to control weeds in a field and then planting corn plantscomprising event MON 87411 in the field. Such application of glyphosateherbicide would be pre-planting, i.e., before MON 87411 containing seedis planted, and could be done any time pre-planting including, but notlimited to, about 14 days pre-planting to about 1 day pre-planting. Theinvention also provides a method for producing corn seed essentiallyfree of weed seeds by planting seeds of a glyphosate tolerant corn plantcomprising MON 87411 in a field, applying a post-emergence effectivedose of glyphosate herbicide sufficient to kill the weed to the field,and harvesting seed from the field. A herbicidally effective dose ofglyphosate for use in the field should consist of a range from about0.125 pounds per acre to about 6.4 pounds per acre of glyphosate over agrowing season. In one embodiment, a total of about 1.5 pounds per acreof glyphosate is applied over a growing season. Multiple applications ofglyphosate may be used over a growing season, for example, twoapplications (such as a pre-planting application and a post-emergenceapplication or a pre-emergence application and a post-emergenceapplication) or three applications (such as a pre-planting application,a pre-emergence application, and a post-emergence application).

Methods for producing an insect and herbicide tolerant corn plantcomprising the DNA sequences specific and unique to event MON 87411 ofthe invention are provided.

Transgenic plants used in these methods may be homozygous orheterozygous for the transgene. Progeny plants produced by these methodsmay be varietal or hybrid plants; may be grown from seeds produced by acorn event MON 87411 containing plant and/or from seeds produced by aplant fertilized with pollen from a corn event MON 87411 containingplant; and may be homozygous or heterozygous for the transgene. Progenyplants may be subsequently self-pollinated to generate a true breedingline of plants, i.e., plants homozygous for the transgene, oralternatively may be outcrossed, e.g., bred with another unrelatedplant, to produce a varietal or a hybrid seed or plant.

Methods of detecting the presence of DNA derived from a corn cell,tissue, seed, or plant comprising corn event MON 87411 in a sample areprovided. One method consists of (i) extracting a DNA sample from atleast one corn cell, tissue, seed, or plant, (ii) contacting the DNAsample with at least one primer that is capable of producing DNAsequence specific to event MON 87411 DNA under conditions appropriatefor DNA sequencing, (iii) performing a DNA sequencing reaction, and then(iv) confirming that the nucleotide sequence comprises a nucleotidesequence specific for event MON 87411, or the construct comprisedtherein, such as one selected from the group consisting of SEQ ID NO:1,SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ IDNO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, SEQ ID NO:12, SEQ IDNO:14, SEQ ID NO:16, SEQ ID NO:21, SEQ ID NO:25, SEQ ID NO: 41, SEQ IDNO: 42, SEQ ID NO: 43, SEQ ID NO: 44, SEQ ID NO: 45, SEQ ID NO: 49, SEQID NO: 50, SEQ ID NO: 51, and SEQ ID NO: 52. Another method consists of(i) extracting a DNA sample from at least one corn cell, tissue, seed,or plant, (ii) contacting the DNA sample with a primer pair that iscapable of producing an amplicon from event MON 87411 DNA underconditions appropriate for DNA amplification, (iii) performing a DNAamplification reaction, and then (iv) detecting the amplicon moleculeand/or confirming that the nucleotide sequence of the amplicon comprisesa nucleotide sequence specific for event MON 87411, such as one selectedfrom the group consisting of SEQ ID NO:21 and SEQ ID NO:25. The ampliconshould be one that is specific for event MON 87411, such as an ampliconthat comprises SEQ ID NO:21 or SEQ ID NO:25. The detection of anucleotide sequence specific for event MON 87411 in the amplicon isdeterminative and/or diagnostic for the presence of the corn event MON87411 specific DNA in the sample. An example of a primer pair that iscapable of producing an amplicon from event MON 87411 DNA underconditions appropriate for DNA amplification is provided as SEQ IDNO:18, SEQ ID NO:24, SEQ ID NO:20, and SEQ ID NO:22. Other primer pairsmay be readily designed by one of skill in the art and would produce anamplicon comprising SEQ ID NO:21 or SEQ ID NO:25, wherein such a primerpair comprises at least one primer within the genomic region flankingthe insert and a second primer within the insert. Another method ofdetecting the presence of DNA derived from a corn cell, tissue, seed, orplant comprising corn event MON 87411 in a sample consists of (i)extracting a DNA sample from at least one corn cell, tissue, seed, orplant, (ii) contacting the DNA sample with a DNA probe specific forevent MON 87411 DNA, (iii) allowing the probe and the DNA sample tohybridize under stringent hybridization conditions, and then (iv)detecting hybridization between the probe and the target DNA sample. Anexample of the sequence a DNA probe that is specific for event MON 87411DNA is provided as SEQ ID NO:19 or SEQ ID NO:23. Other probes may bereadily designed by one of skill in the art and would comprise at leastone fragment of genomic DNA flanking the insert and at least onefragment of insert DNA, such as sequences provided in, but not limitedto, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQID NO:8, SEQ ID NO:9, SEQ ID NO:10, SEQ ID NO:21, and SEQ ID NO:25.Detection of probe hybridization to the DNA sample is diagnostic for thepresence of corn event MON 87411 specific DNA in the sample. Absence ofhybridization is alternatively diagnostic of the absence of corn eventMON 87411 specific DNA in the sample.

DNA detection kits are provided that are useful for the identificationof corn event MON 87411 DNA in a sample and can also be applied tomethods for breeding corn plants containing the appropriate event DNA.Such kits contain DNA primers and/or probes comprising fragments of SEQID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ IDNO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, SEQ ID NO:12,SEQ ID NO:14, SEQ ID NO:16, SEQ ID NO:21, SEQ ID NO:25, SEQ ID NO: 41,SEQ ID NO: 42, SEQ ID NO: 43, SEQ ID NO: 44, SEQ ID NO: 45, SEQ ID NO:49, SEQ ID NO: 50, SEQ ID NO: 51, and SEQ ID NO: 52. One example of sucha kit comprises at least one DNA molecule of sufficient length ofcontiguous nucleotides of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ IDNO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9,SEQ ID NO:10, SEQ ID NO:12, SEQ ID NO:14, SEQ ID NO:16, SEQ ID NO:21,SEQ ID NO:25, SEQ ID NO: 41, SEQ ID NO: 42, SEQ ID NO: 43, SEQ ID NO:44, SEQ ID NO: 45, SEQ ID NO: 49, SEQ ID NO: 50, SEQ ID NO: 51, and SEQID NO: 52 to function as a DNA probe useful for detecting the presenceand/or absence of DNA derived from transgenic corn plants comprisingevent MON 87411 in a sample. The DNA derived from transgenic corn plantscomprising event MON 87411 would comprise a DNA molecule having at leastone sequence selected from the group consisting of SEQ ID NO:1, SEQ IDNO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7,SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, SEQ ID NO:12, SEQ ID NO:14, SEQID NO:16, SEQ ID NO:21, SEQ ID NO:25, SEQ ID NO: 41, SEQ ID NO: 42, SEQID NO: 43, SEQ ID NO: 44, SEQ ID NO: 45, SEQ ID NO: 49, SEQ ID NO: 50,SEQ ID NO: 51, and SEQ ID NO: 52. A DNA molecule sufficient for use as aDNA probe is provided that is useful for determining, detecting, ordiagnosing the presence and/or absence of corn event MON 87411 DNA in asample is provided as SEQ ID NO:19 and SEQ ID NO:23. Other probes may bereadily designed by one of skill in the art and should comprise asufficient number of contiguous nucleic acids, including at least 15, atleast 16, at least 17, at least 18, at least 19, at least 20, at least21, at least 22, at least 23, at least 24, at least 25, at least 26, atleast 27, at least 28, at least 29, at least 30, at least 31, at least32, at least 33, at least 34, at least 35, at least 36, at least 37, atleast 38, at least 39, or at least 40 contiguous nucleotides of SEQ IDNO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6,SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, SEQ ID NO:12, SEQID NO:14, SEQ ID NO:16, SEQ ID NO:21, SEQ ID NO:25, SEQ ID NO: 41, SEQID NO: 42, SEQ ID NO: 43, SEQ ID NO: 44, SEQ ID NO: 45, SEQ ID NO: 49,SEQ ID NO: 50, SEQ ID NO: 51, and SEQ ID NO: 52 and be sufficientlyunique to corn event MON 87411 DNA in order to identify DNA derived fromthe event. Another type of kit comprises a primer pair useful forproducing an amplicon useful for detecting the presence and/or absenceof DNA derived from transgenic corn event MON 87411 in a sample. Such akit would employ a method comprising contacting a target DNA sample witha primer pair as described herein, then performing a nucleic acidamplification reaction sufficient to produce an amplicon comprising aDNA molecule having at least one sequence selected from the groupconsisting of SEQ ID NO:21 and SEQ ID NO:25, and then detecting thepresence and/or absence of the amplicon. Such a method may also includesequencing the amplicon or a fragment thereof, which would bedeterminative of, i.e. diagnostic for, the presence of the corn eventMON 87411 specific DNA in the target DNA sample. Other primer pairs maybe readily designed by one of skill in the art and should comprise asufficient number of contiguous nucleic acids, including at least 15, atleast 16, at least 17, at least 18, at least 19, at least 20, at least21, at least 22, at least 23, at least 24, at least 25, at least 26, atleast 27, at least 28, at least 29, or at least 30 contiguousnucleotides of sequences provided in, but not limited to SEQ ID NO:1,SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ IDNO:8, SEQ ID NO:9, or SEQ ID NO:10, SEQ ID NO:12, SEQ ID NO:14, SEQ IDNO:16, SEQ ID NO: 41, SEQ ID NO: 42, SEQ ID NO: 43, SEQ ID NO: 44, SEQID NO: 45, SEQ ID NO: 49, SEQ ID NO: 50, SEQ ID NO: 51, and SEQ ID NO:52 and be sufficiently unique to corn event MON 87411 DNA in order toidentify DNA derived from the event.

The kits and detection methods of the invention are useful for, amongother things, identifying corn event MON 87411, selecting plantvarieties or hybrids comprising corn event MON 87411, detecting thepresence of DNA derived from the transgenic corn plants comprising eventMON 87411 in a sample, and monitoring samples for the presence and/orabsence of corn plants comprising event MON 87411 or plant parts derivedfrom corn plants comprising event MON 87411.

The sequence of the heterologous DNA insert, junction sequences, orflanking sequences from corn event MON 87411 can be verified (andcorrected if necessary) by amplifying such sequences from the eventusing primers derived from the sequences provided herein followed bystandard DNA sequencing of the amplicon or of the cloned DNA.

The following examples are included to demonstrate examples of certainpreferred embodiments of the invention. It should be appreciated bythose of skill in the art that the techniques disclosed in the examplesthat follow represent approaches the inventors have found function wellin the practice of the invention, and thus can be considered toconstitute examples of preferred modes for its practice. However, thoseof skill in the art should, in light of the present disclosure,appreciate that many changes can be made in the specific embodimentsthat are disclosed and still obtain a like or similar result withoutdeparting from the spirit and scope of the invention.

Deposit Information

A deposit of a representative sample of corn seed comprising event MON87411 has been made on Mar. 14, 2012 according to the Budapest Treatywith the American Type Culture Collection (ATCC) having an address at10801 University Boulevard, Manassas, Virginia USA, Zip Code 20110, andassigned ATCC Accession No. PTA-12669. Access to the deposits will beavailable during the pendency of the application to the Commissioner ofPatents and Trademarks and persons determined by the Commissioner to beentitled thereto upon request. Upon issuance of the patent, allrestrictions upon availability to the public will be irrevocablyremoved. The deposit will be maintained in the depository for a periodof 30 years, or 5 years after the last request, or for the effectivelife of the patent, whichever is longer, and will be replaced asnecessary during that period.

EXAMPLES Example 1

This example describes the design and selection of a constructdesignated 417 and the engineering and evaluation of different DNAconstructs. Table 1 tabulates these DNA constructs by test criteria andresults.

DNA constructs were engineered to express an RNA-basedplant-incorporated protectant (PIP) in corn, targeting Western cornrootworm (WCR). Variations of the RNA transcript were tested fordifferent target genes of WCR (Group 1), different lengths of RNA (Group2), with or without neutral RNA carrier (Group 2), different secondarystructures (Group 4), and different target segments of Dv_Snf7o (Groups2 and 3). Variations on multiple transgenes were also tested, e.g., theRNA transcript+a WCR-active protein (Groups 3 and 5), and two RNAtranscripts targeting two WCR targets (Groups 1 and 4). Variations onthe number and configuration of expression cassettes and elements usedwere also tested (all groups).

TABLE 1 Forty-five DNA constructs were stably transformed into cornplants. Progeny plants from multiple transformation events per DNAconstruct were evaluated. Construct Group Criteria and Results 043 1Tested inhibition of WCR activity on plants expressing 043 vectorstacked combinations of RNA segments targeting 059 transcripts of 4different WCR endogenous genes. WCR activity was inhibited on plantsexpressing an RNA segment targeting the Dv_Snf7o gene transcript. 503 2Tested inhibition of WCR activity on plants expressing 475 various sizesof RNA segments targeting the Dv_Snf7o 970 gene transcript (from a27-mer up to a 429-mer) 474 engineered to express as an inverted-repeatRNA (IR). 477 Also tested a 150-mer neutral IR carrier that was 306embedded with and without a 27-mer targeting Dv_Snf7o. 476 Optimal WCRactivity was observed on plants expressing 713 Dv_Snf7o target segmentsequal or longer than 100 base pairs in length. 868 3 Tested inhibitionof WCR activity on plants expressing: 870 (a) a 240-mer Dv_Snf7o IR, and(b) a pair of proteins 871 TIC809 and TIC810 having WCR inhibitoryactivity; both 875 under one expression cassette in one DNA construct.310 Tested inhibition of WCR activity on plants expressing: 311 (a) the240-mer Dv_Snf7o IR, and (b) the pair of proteins 330 TIC809 and TIC810having WCR inhibitory activity; each 331 independently- andoperably-linked to separate expression 950 cassettes in one DNAconstruct. 890 Tested these IR + protein combinations using different867 combinations of different promoters and expression 946 cassetteconfigurations. 878 In-planta expression of the 240-mer Dv_Snf7o IR 823inhibited WCR activity on such plants, with or without 879 expression ofthe TIC809 and TIC810 protein pair. 880 401 354 4 Tested progeny plantsof a hybrid cross between plants 253 containing events harboring DNAconstruct #503 (a 429- 254 mer Dv_Snf7o IR) and plants comprising eventMON 255 88017 (Cry3Bb). 256 Tested inhibition of WCR activity on plantsexpressing a 892 150- or 240-mer Dv_Snf7o IR. 365 Tested inhibition ofWCR activity on plants expressing: (a) Dv_Snf7o IR, and (b) vATPase AIR. Tested IR versus non-IR secondary RNA structures for suppressingDv_Snf7o, vATPase A, and the combination. In-planta Expression of the240-mer Dv_Snf7o IR inhibited WCR activity, with or without expressionof the vATPase A RNA segment. WCR inhibition was better in-planta whenDv_Snf7o IR was expressed together with Cry3Bb, when compared toexpressing Dv_Snf7o IR alone or Cry3Bb alone. 416 5 Tested inhibition ofWCR activity on plants expressing 417 both (a) the 240-mer Dv_Snf7o IR,and (b) the Cry3Bb 418 protein having Diabrotica virgifera pesticidalactivity; 419 each transgene in separate expression cassettes in a DNA423 construct. 402 Tested ten DNA constructs having combinations of 403different promoters, and combinations of different 404 expressioncassette configurations. 405 DNA construct #417 was selected. 406

Using the DNA constructs of Group 2 as an example, 7 DNA constructs wereengineered to test the targeting of various lengths of Dv_Snf7o (from 27up to 429 nt in length). Each DNA construct was produced, plant cellstransformed, plants obtained, and inbreds evaluated in growth chamberefficacy bioassays. Results showed a correlation between length ofinverted repeat RNA (IR) and WCR activity (Table 2, columns (B) and(H)).

TABLE 2 Correlation between length of IR and WCR-activity. (F) (G) (B)No. of R₀ No. of (A) Dv_Snf7o (C) (D) plants events (H) DNA RNA No. ofNo. of (E) expected to advanced for WCR- Construct segment embryosembryos No. of R₀ harbor a multi-plant activity on No. length (nt)transformed w/shoots plants to soil single event testing plants? 503 4292085  433  308  233  78 +++++ 475 150 230 57 45 39 23 +++++ 970  27† 22079 47 44 21 ++ 474  27 230 81 51 49 23 − 477  50 220 50 36 31 23 ++ 306 75 230 37 27 18 15 ++ 476 100 220 53 40 33 22 +++++

Column (B) displays the variable lengths of Dv_Snf7o target RNAengineered to express as an inverted repeat RNA (IR) secondary structurein corn plants. Column (C) displays the number of corn embryos that weretransformed. Column (D) displays the number of corn embryos thatdeveloped shoots. Column (E) displays the number of regenerated cornplants (designated as generation R0) viable on soil. Column (F) displaysthe number of R0 plants expected to harbor a single copy of insert DNAin the transformation event. Column (G) displays the number of R0 plantsthat were expected to harbor a single transformation event, and thatproduced enough seed for multi-plant growth chamber bioassay. Column (H)displays the results of plant growth chamber studies designed toevaluate WCR-activity. “+++++” indicates average RDR was less than 0.5RDR. “++” indicates average RDR was between 0.5 RDR and 2.0 RDR. “−”means average RDR was about 2.0 RDR, which was comparable to negativecontrols in growth chamber efficacy studies.

† the same 27-mer as in DNA construct #474 but embedded in a neutral150-mer IR. To evaluate WCR activity on plants grown in growth chambers,6 to 8 plants for each of 10-20 events per construct were grown in peatpots. Plants were tested for the presence of the insert DNA and forexpression of the transgene(s) in both leaf and root tissues. Plantsconfirmed to have expression of the transgene were then transplantedinto larger pots infested with WCR eggs. Non-transgenic corn lines LH59and LH244 were included as negative controls. Plants containing eventMON 88017 (expressing Cry3Bb) were included as positive controls. Rootdamage of the growing corn plants was assessed after 4 weeks. Rootdamage ratings (RDR) were assessed on a three-point scale, with 0 RDRhaving no root damage and 3 RDR having maximum root damage.

Study results guided the design of the DNA constructs of Group 5 tocontain (a) an expression cassette for a 240-mer Dv_Snf7o IR, and (b) anexpression cassette for a Cry3Bb protein (FIG. 2 ). The 240-mer Dv_Snf7oIR was selected because (a) plants expressing the identical 240-merDv_Snf7o IR were repeatedly successful in inhibiting CRW activity(Groups 2-4), (b) segments larger than 100 nt in length decrease theprobability of development of WCR resistance, and (c) segments largerthan 240 nt would make it more difficult to transfer intact into thecorn genome. The DNA constructs were designed to test differentregulatory genetic elements in each expression cassette and differentconfigurations of each expression cassette in the DNA construct. DNAconstructs of Group 5 also included constructs with and withoutglyphosate tolerance expression cassettes; and a control construct fromgroup 3 that expressed only the 240-mer Dv_Snf7o IR. Each DNA constructwas designed, plant cells transformed, plants obtained, and inbredsevaluated in growth chamber efficacy bioassays (Table 3 (C) through(H)).

TABLE 3 Plant production numbers from transformation of Group 5 DNAconstructs. (F) (G) (H) (D) (E) Number of Number of Inbred and (A) (B)(C) Number Number R₀ plants R₀ events hybrid DNA DNA Number of of of R₀expected to advanced to progeny Construct construct embryos embryosplants to harbor a growth plant No. composition transformed w/shootssoil single event chamber performance (1) 416 Dv_Snf7o 820 72 72 42 27+++++ (2) 417 IR + 521 212 94 71 44 +++++ (3) 418 Cry3Bb + 588 79 65 4428 +++++ (4) 419 EPSPS 651 106 95 68 43 ++++ (5) 423 754 93 84 66 41++++ (6) 402 786 84 84 58 43 ++++ (7) 403 714 199 84 46 40 ++++ (8) 404740 50 50 34 29 ++++ (9) 405 Dv_Snf7o 21663 1586 1586 86 58 +++ (10) 406 IR + Cry3Bb 21965 1539 1539 170 112 ++++ (11)  890 Dv_Snf7o 3996 656394 235 136 +++ IR Column (A) lists the DNA constructs tested in stage 5(also see FIG. 2 for breakdown of the genetic elements). Column (B)displays the combination of transgene. Column (C) displays the number ofcorn embryos that were transformed. Column (D) displays the number ofcorn embryos that developed shoots. Column (E) displays the number ofregenerated corn plants (designated as generation R0) viable on soil.Column (F) displays the number of R0 plants expected to harbor a singletransformation event. Column (G) displays the number of R0 plantsexpected to harbor a single transformation event, and that producedenough seed for subsequent multi-plant testing. Column (H) summarizesthe performance of plants infested with WCR (See following paragraph fordetails).

As shown in Table 3, column (H), “+++++” describes DNA constructs thaton average provided the highest sustained gene expression to transgenicplants throughout their development, most WCR inhibition duringdevelopment, and most WCR inhibition in self-fertilized andcross-hybridized generations. “++++” describes DNA constructs that onaverage provided WCR inhibition to transgenic plants but lower geneexpression when compared to the “+++++” plants. “+++” describes DNAconstructs that on average provided lower WCR inhibition to transgenicplants when compared to the “++++” and “+++++” plants. Therefore, DNAconstruct #417 was advanced for further analysis. This construct hassixteen genetic elements organized into three expression cassettes fromthe Left Border (LB) through to the Right Border (RB). The construct isshown in FIG. 2 and the sequence given in SEQ ID NO:26. The vectorcomponents are as follows:

[1] LB: Corresponds to the reverse complement of positions 1 through 442of SEQ ID NO:26. This element represents the octopine Left bordersequence from Agrobacterium tumefaciens.

[2] Ps.RbcS2-E9 3′ UTR: Corresponds to the reverse complement ofpositions 486 through 1118 of SEQ ID NO:26. Represents 3′ untranslatedregion (UTR) from the ribulose 1,5-bisphosphate carboxylase smallsubunit E9 (rbcS-E9) gene transcript from Pisum sativum (pea).

[3] 240-mer Dv_Snf7o inverted repeat gene: Corresponds to the reversecomplement of positions 1148 through 1777 of SEQ ID NO:26. This genetranscribes RNA containing two 240-mer ribonucleotide segments thatalign identically to each other in reverse complement fashion, separatedby a neutral segment of 150 ribonucleotides, and forming an invertedrepeat RNA (IR). The sequence of the 240-bp segment aligns to a WCR geneorthologous to yeast Snf7.

[4] Corn DnaK intron: Corresponds to the reverse complement of positions1814 through 2617 of SEQ ID NO:26. This element consists of 10nucleotides of exon 1, intron 1, and 11 nucleotides of exon 2 from theheat shock protein 70 gene from Zea mays (corn). The 11 nucleotides ofexon 2 were modified to remove an initiating methionine residue.

[5] CaMV 35S leader: Corresponds to the reverse complement of positions2618-2626 of SEQ ID NO:26. Represents the 5′ untranslated region (UTR)from the 35S RNA transcript of the Cauliflower mosaic virus (CaMV)beginning at the +1 position of the mRNA transcriptional start of thegene.

[6] eCaMV 35S promoter: Corresponds to the reverse complement ofpositions 2627-3238 of SEQ ID NO:26. Represents the promoter of 35S RNAfrom Cauliflower mosaic virus (CaMV) containing a duplication of the −90to −350 region.

[7] Corn PIIG promoter: Corresponds to positions 3265-4213 of SEQ IDNO:26. This genetic element represents the promoter of the physicalimpedance induced protein (PIIG) gene from Zea mays.

[8] Wheat Lhcb1 leader: Corresponds to positions 4220-4280 of SEQ IDNO:26. This genetic element represents the 5′ untranslated region (UTR)of the light harvesting complex b1 (Lhcb1) gene from Triticum aestivum(wheat).

[9] Rice Act1 intron: Corresponds to positions 4297-4776 of SEQ IDNO:26. Consists of a contiguous sequence of 12 nucleotides of exon 1,intron 1, and 7 nucleotides of exon 2 from the Actin 1 (Act1) gene ofOryza sativa (rice).

[10] Cry3Bb ORF: Corresponds to positions 4786-6747 of SEQ ID NO:26.Represents the coding region of a non-naturally occurring pesticidalCry3B protein engineered to exhibit modifications H231R, S311L, N313T,E317K, and Q349R as compared to the native Bt Cry3Bb protein encodinggene. The nucleotide sequence aligns to the cry3Bb gene sequencecontained in event MON 88017.

[11] Wheat Hsp17 3′ UTR: Corresponds to positions 6767-6976 of SEQ IDNO:26. This genetic element represents the 3′ UTR of the heat shockprotein 17 (HSP17) gene from Triticum aestivum (wheat).

[12] Rice TubA (promoter, leader, intron): Corresponds to positions7025-9205 of SEQ ID NO:26. Represents the contiguous promoter, leader,intron, and 4 nucleotides of exon 2 from the alpha tubulin gene (TubA-3)of Oryza sativa (rice).

[13] CTP: Corresponds to positions 9210-9437 of SEQ ID NO:26. Representsengineered coding region encoding the N-terminal CTP from5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) from A. thaliana.This element differs from the native gene (GenBank Accession No. X06613)at the last GAG codon (glutamic acid) by modification to TGC (cysteine).

[14] CP4 EPSPS: Corresponds to positions 9438-10805 of SEQ ID NO:26.Represents engineered coding region of the EPSPS from Agrobacterium CP4.Differs from the native Agrobacterium gene at the second codon bymodification from encoding serine to CTT (leucine) and four silentsubstitutions.

[15] Rice TubA 3′ UTR: Corresponds to positions 10813-11394 of SEQ IDNO:26. Represents the 3′ untranslated region (UTR) of an alpha tubulingene (TubA-3) from Oryza sativa (rice).

[16] RB: Corresponds to positions 11413-11743 of SEQ ID NO:26.Represents nopaline right border sequence from A. tumefaciens.

Example 2

This example describes the transformation and selection of event MON87411 from among a plurality of transgenic events.

Embryos were excised from kernels of corn line LH244, and inoculatedwith recombinant Agrobacterium harboring DNA construct #417. Co-culturedembryos were transferred onto selection and growth media to generatetransgenic callus tissue with developing shoots. Developing shoots weretransferred to rooting medium for development into plantlets. Plantletswere regenerated into whole R₀ plants in soil. R₀ plants recovered thisway were screened for a single copy of introduced construct DNA. Asshown in Table 3, putative single-copy events were provided in 71 uniqueR₀ transformants. Each R₀ transformant was placed under nurseryconditions to produce progeny R₁ seed. Forth-four events were advanced.At least 8 R₁ seeds produced by each of the 44 R₀ plants were planted insoil and R₁ plants were grown to produce R₂ seed. A single R₁ plant perevent was selected to continue each line containing each separate event,and seed from the single R₁ plant was bulked for subsequent testing by(a) self-fertilization (R_(3, 4, . . . , N)), and (b)cross-fertilization with other corn lines, e.g., corn line 93IDI3.Plants representing events from transformation of DNA construct #890(row 11 of Table 3) were also regenerated to serve as comparativecontrols for subsequent field trials described below and in thisexample.

Of the 44 events, 25 events were chosen to go forward based on aphenotype including Cry3Bb expression. The R₁ plants representing these25 events were further evaluated for WCR inhibition in growth chamberefficacy methods described in Example 1, and for copy-number of multiplegenetic elements of the insert DNA. Seventeen events out of the 25events were taken forward, as four events exhibited more than one copyof the Ps.RbcS2-E9 3′ UTR genetic element, and R₁ plants representing 4other events exhibited root damage ratings greater than 0.8 RDR.

Progeny plants comprising the remaining 17 events, i.e., “A”, MON 87411,and “C” through “Q”, were further analyzed in parallel for molecular andfor in-field performance (see Tables 4 and 5).

TABLE 4 Molecular analysis of 17 transgenic corn events harboring insertDNA from DNA transformation vector #417. (D) (E) (B) Above Above SingleInsert threshold threshold IR (F) (G) (A) and Single Cry3Bb Dv_Snf7oNeutral Expected Backbone Copy- (C) protein dsRNA insertion transcriptEvent absent number Intact insert expression expression site sizeA + + + + + + + MON 87411 + + + + + + + C + + + + + + + D + + − + + + +E + + + + + − NA F + + + + + − NA G + + + + + − NA H + + NA NA NA NA NAI + + NA NA NA NA NA J + + NA NA NA NA NA K + − NA NA NA NA NA L − − NANA NA NA NA M − + NA NA NA NA NA N − − NA NA NA NA NA O − + NA NA NA NANA P − − NA NA NA NA NA Q − − NA NA NA NA NA “−” indicates that theevent did not meet the molecular criteria of the corresponding molecularanalysis. “+” indicates that the event met the molecular criteria of thecorrespoding molecular analysis. “NA” indicates that the data was notavailable.

Events were screened for backbone DNA segments of the Agrobacteriumtransformation vector and for single copy-number of all portions of theintended insert DNA (Table 4, Columns (A) and (B)). Seven events (MON87411, A, C, D, E, F, and G) were analyzed for sequence of the insertedDNA which identical to the transformation vector #417, with theexception of nick site variations at the agrobacterium left and rightborders that occur during Agro-mediated insertion, event D failed thissequence analysis (Table 4, Column (C)). These 7 events were alsoevaluated for sustained plant expression of Cry3Bb protein and Dv_Snf7oIR RNA throughout plant development and several generations, and all 7events met the passing criteria for sustained plant expression (Table 4,Column (D)). Each of the 7 events were analyzed for genomic insertionsite characteristics (i.e., neutral insertion site), such as DNAdisplacement, duplications and repetitiveness, proximity to anendogenous gene, interruption of an endogenous gene, and proximity toQTLs and biotech traits, events E, F, and G failed this analysis (Table4, Column (F)). Northern blots were performed on plant tissue containingevents MON 87411, A, C, and D to determine if the expected sizes of thetwo RNA transcript encoding Cry3Bb, or producing the Dv_Snf7o IR RNAwere present in RNA from the events, and all events evaluated passedthis criteria (Table 4, Column (G)).

These 17 events were evaluated in agronomic, insect efficacy andglyphosate tolerance efficacy field trials, the results are summarizedin Table 5. The column headers of Table 5 describe the type of fieldtrial (“Agronomics”, “Insect:”, or “Glyphosate”), the controls to whichthe events were being compared/contrasted are listed, and the geneticinbred used to generate event hybrid is also listed. The field trialssummarized in columns (A) through (C) were planted one calendar yearbefore the field trials summarized in columns (D) through (H), and twoyears before the field trials summarized in column (I).

TABLE 5 Results from Agronomic, Insect efficacy, and glyphosate efficacyfield trials of events generated with transformation vector #417. (F)(G) (H) (A) (B) (C) (D) Glyphosate Glyphosate Insect (I) Type of fieldtrial Agronomics Agronomics Insect Efficacy Agronomics Efficacy EfficacyEfficacy Agronomics Controls used as LH244, LH244 × 93IDI3, LH244,LH244, MON MON MON 88017, LH244, comparison #890 #890 MON 88017, #890MON 88017 88017 88017 #890 #890

R3 inbred R3 inbred R2 inbred × 93IDI3 R5 inbred R5 inbred R4 inbred ×R4 inbred × R5 inbred MON 89034 MON 89034 Test Event A = = <0.10 RDR = == ~0.10 RDR − MON 87411 = = NA = = = ~0.10 RDR = C = = ~0.10 RDR = − NANA NA D = = ~0.10 RDR + = = ~0.20 RDR NA E = = NA + = = ~0.15 RDR = F == NA + − NA NA NA G = = NA + = = ~0.15 RDR = H‡ − = ~0.10 RDR NA NA NANA NA I‡ − = NA NA NA NA NA NA J† = = NA NA NA NA NA NA K − = ~0.15 RDR= NA NA NA NA L = = NA = NA NA NA NA M = = ~0.20 RDR + NA NA NA NA N − =NA − NA NA NA NA O = = NA NA NA NA NA NA P − = NA NA NA NA NA NA Q = =NA NA NA NA NA NA

Events were compared to control(s) in each field trial. Data for eachfield trial were averaged by replicate plots over multiple locations.LH244 is the control for the transformation line. The DNA vector “#890”was used to produce events expressing only the 240-mer Dv_Snf7o IR. Thecommercial event, MON 88017, which provides coleopteran resistance andglyphosate tolerance to corn plants was used as a control. “R_(N)inbred” specifies the N^(th) generation progeny. Hybrid events evaluatedin the field trials were grown from seed harvested from a cross with oneparent from the event under evaluation (MON 87411, or A through Q), andone parent as indicated in Table 5 (Column C, G, or H). Specifically, inTable 5, column R2 inbred X 93IDI3 specifies that an R2 inbred of theevent under evaluation was crossed with inbred corn line 93IDI3 to makethe hybrid seed. Similarly, in Table 5, columns G and H, R4 inbred X MON89034 specifies that an R4 inbred progeny of the event under evaluationwas crossed with a plant containing event MON 89034 to make the hybridseed. “NA” indicates that data for this test event was not available.“=” represents trait equivalency compared to controls. “−” represents atrait hit compared to controls. “+” represents an increase inperformance compared to controls. “RDR” is root damage rating. “‡”represents that contemporaneous greenhouse studies showed that theapplicable event exhibited phenotypic off-types in plants grown in thenursery. “†” represents that contemporaneous greenhouse studies showedthat the applicable event did not provide WCR efficacy.

Agronomic field trials were conducted at multiple North American andSouth American locations, the results were averaged across alllocations. as summarized Table 5, columns A, B, D, and I. For theseagronomic field trials, corn kernels were planted in a randomizedcomplete block (RCB) design in triplicate plots per event per location.Each replicate plot consisted of 100 kernels. Trial maintenance wasdesigned to optimize grain production and eliminate natural WCR pressureOne or more of the following standard agronomic field trial ratings werecollected: degree units to 50% shed (GDU), Breeder's score (BR),seedling vigor (SDV), stalk lodging (STLC), root lodging (RTLC), earheight of mature plants (EHT), plant height of mature plants (PHT),grain moisture (MST), and grain test weight (TWT), phenotypic off-types,and grain yield. Both inbred and hybrid events were evaluated and theresults are summarized in Table 5, columns A, B, D, and I. Appropriatecontrols were included in triplicate plots per control per location. Theratings were averaged by plot across all locations. Data were subjectedto an analysis of variance and means separated at the least significantdifference at the 5% probability level (LSD (0.05)).

Results of insect efficacy field trials that included analyses for WCRdamage averaged across multiple North American locations are summarizedin Table 5, columns C and H. For these efficacy field trials, cornkernels were planted in a RCB design in triplicate plots per event perlocation; each replicate plot consisted of 25 kernels. Test events werepresented in hybrid plants. Appropriate controls were included intriplicate plots per control per location. When plots of corn reachedtheir V2 growth stage, 5 plants per plot were infested with WCR eggs ata rate of 3,330 eggs per plant. During the V10 growth stage, the rootsof the 5 infested plants per plot were dug up, washed, and evaluated forfeeding damage based on a root damage rating (RDR) of 0 to 3, with 0 RDRhaving no root damage and 3 RDR having maximum root damage. RDRs fortest events and control plants were averaged by plant across all plotsin all locations. Negative control plants of each insect efficacy fieldtrial exhibited respective average RDRs of 1.7 and 1.5 RDR. Commercialchecks of each insect efficacy field trial exhibited respective averageRDRs of 0.25 and 0.20 RDR. Plants containing events from DNA construct#890 exhibited a range of RDRs from about 0.35 to 0.50 RDR. Events fromDNA Construct #417 consistently provided plants with average RDR scoresless than the economic injury threshold of 0.25 RDR.

Results of efficacy field trials evaluating vegetative tolerance toglyphosate herbicide treatments were conducted across multiple NorthAmerican locations and are summarized in Table 5, columns F and G. Forthese efficacy field trials, the glyphosate application regimen used forthe specific trial is presented in Table 6 (corresponding to Table 5,column F) and Table 7 (corresponding to Table 5, column G).

TABLE 6 Herbicide Field Trial Treatments. Treatment Rate (lbs ae/A)Schedule (by plant stage) Glyphosate 1.5 V2 Glyphosate 1.5, 0.75, 0.75V2, V8, V10 Glyphosate 1.5, 1.125, 1.125 V2, V8, V10 “lbs ae” indicatespound acid equivalent. “A” indicates acre.

TABLE 7 Herbicide Field Trial Treatments. Treatment Rate (lbs ae/A)Schedule (by plant stage) Untreated 0.0 n/a Glyphosate 1.5, 1.5 V4, V8Glyphosate 3.0, 3.0 V4, V8 Glyphosate 4.5, 4.5 V4, V8 “lbs ae” indicatespound acid equivalent. “A” indicates acre.

Each plot of 100 plants was rated for crop injury 7-10 days after thelast spray of each treatment. Crop injury ratings included chlorosis,malformation, and average lower plant height, all of which indicatelower tolerance to the glyphosate herbicide. Each plot was also ratedfor PHT, EHT, days to 50% pollen shed (D50P), days to 50% silk emergence(D50S), TWT, MST, and yield. Events were provided as inbred plants andhybrid plants and compared to event MON 88017. Events “A”, MON 87411,“D”, “E”, and “G” were equivalent to event MON 88017 relative to cropinjury, PHT, EHT, D50P, D50S, TWT, MST, and yield ratings. Based onthese results coupled with the significant RDR advantage of eventMON87411 compared to other events and to the commercial MON88017 event,event MON 87411 was selected.

Example 3

This example describes the molecular characterization of event MON87411. A sample of Leaf tissue was sampled from an (R₀) MON87411 plant.Sequencing of the genomic DNA corresponding to the transgenic insertionsite in event MON 87411 was obtained and no differences were observedcompared to the sequence in the transformation vector corresponding tovector #417.

The flanking sequences were mapped to corn genome reference sequences,including the maize B73 reference genome (Ref B73). Event MON 87411 wasdetermined to be physically located on chromosome 9. The flankingsequence ending at the left flank/insert DNA junction corresponds toposition ZM_B73_CR09:39261797. The flanking sequence ending at the rightflank/insert DNA junction corresponds to position ZM_B73_CR09:39261915.The flanking sequences for event MON 87411 were analyzed for genomeduplications, repeats, and endogenous genes. None were detected.

The sequence analysis of the inserted DNA in event MON 87411 confirmedthat only 263 nucleotides of the Agrobacterium left border (arbitrarilyset as the 5′ end of the insert), and only 15 nucleotides of theAgrobacterium right border (arbitrarily set as the 3′ end of the insert)were retained in the inserted DNA at the genomic insertion site of eventMON 87411.

A comparative analysis of the genomic sequence flanking the inserted DNAof event MON 87411 and the corresponding genomic region of the site ofinsertion in the wild-type allele from LH244 was conducted. Thisanalysis determined that a 118 base pair segment of LH244 genomic DNAwas displaced by the inserted DNA of the transformation vector #417 inthe process of generating event MON 87411.

Example 4

This example describes methods which are useful in identifying thepresence of DNA derived from event MON 87411 in a corn sample. A pair ofprimers and a probe were designed for the purpose of identifying theunique junction formed between the genomic DNA and the arbitrarilyassigned 5′ end of the inserted DNA of event MON 87411 (i.e., the leftjunction) and encompassed in SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:4, SEQID NO:5, SEQ ID NO:6, SEQ ID NO:7, or SEQ ID NO:21. The sequence of theoligonucleotide forward primer SQ27011 (SEQ ID NO:18) is identical tothe nucleotide sequence corresponding to positions 462 through 490 ofSEQ ID NO:1 and SEQ ID NO:2, positions 107 through 135 of SEQ ID NO:7,positions 72 through 100 of SEQ ID NO:6, positions 12 through 40 of SEQID NO:5, and positions 1 through 29 of SEQ ID NO:21. The sequence of theoligonucleotide reverse primer SQ9085 (SEQ ID NO:20) is identical to thereverse complement of the nucleotide sequence corresponding to positions516 through 541 of SEQ ID NO:1 and SEQ ID NO:2, positions 161 through186 of SEQ ID NO:7, positions 126 through 151 of SEQ ID NO:6, positions66 through 91 of SEQ ID NO:5, positions 16 through 41 of SEQ ID NO:4,and positions 55 through 80 of SEQ ID NO:21. The sequence of theoligonucleotide probe PB3552 (SEQ ID NO:19) is identical to the reversecomplement of the nucleotide sequence corresponding to positions 502through 515 of SEQ ID NO:1 and SEQ ID NO:2, positions 147 through 160 ofSEQ ID NO:7, positions 112 through 125 of SEQ ID NO:6, positions 52through 65 of SEQ ID NO:5, positions 2 through 15 of SEQ ID NO:4, andpositions 41 through 54 of SEQ ID NO:21. The PCR primers SQ27011 (SEQ IDNO:18) and SQ9085 (SEQ ID NO:20) amplify a 79 nucleotide amplicon of theunique the genomic/insert DNA at the left junction of event MON 87411.This same primer pair with probe PB3552 (SEQ ID NO:19), which has beenfluorescently labeled (i.e., a 6FAM™ fluorescent label), can be used inan Endpoint TaqMan® PCR assay to identify the presence of DNA derivedfrom event MON 87411 in a sample.

A pair of primers and a probe were designed for the purpose ofidentifying the unique junction formed between the genomic DNA and thearbitrarily assigned 3′ end of the inserted DNA of event MON 87411(i.e., the right junction) and encompassed in SEQ ID NO:1, SEQ ID NO:3,SEQ ID NO:4, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, or SEQ ID NO:25.The sequence of the oligonucleotide forward primer SQ27066 (SEQ IDNO:22) is identical to the nucleotide sequence corresponding topositions 11710 through 11728 of SEQ ID NO:1, positions 11210 through11228 of SEQ ID NO:4, positions 45 through 63 of SEQ ID NO:8, SEQ IDNO:9, and SEQ ID NO:10, and positions 1 through 19 of SEQ ID NO:25. Thesequence of the oligonucleotide reverse primer SQ26977 (SEQ ID NO:24) isidentical to the reverse complement of the nucleotide sequencecorresponding to positions 11756 through 11784 of SEQ ID NO:1, positions91 through 117 of SEQ ID NO:8, positions 91 through 119 of SEQ ID NO:9and SEQ ID NO:10, positions 23 through 51 of SEQ ID NO:3, and positions47 through 75 of SEQ ID NO:25. The sequence of the oligonucleotide probePB11300 (SEQ ID NO:23) is identical to the nucleotide sequencecorresponding to positions 11731 through 11755 of SEQ ID NO:1, positions11231 through 11248 of SEQ ID NO:4, positions 66 through 90 of SEQ IDNO:8, SEQ ID NO:9, and SEQ ID NO:10, positions 1 through 22 of SEQ IDNO:3, and positions 22 through 46 of SEQ ID NO:25. The PCR primersSQ27066 (SEQ ID NO:22) and SQ26977 (SEQ ID NO:24) amplify a 75nucleotide amplicon of the unique the genomic/insert DNA at the rightjunction of event MON 87411. This same primer pair with probe PB11300(SEQ ID NO:23), which has been fluorescently labeled (i.e., a 6FAM™fluorescent label), can be used in an Endpoint TaqMan® PCR assay toidentify the presence of DNA derived from event MON 87411 in a sample.

In addition to SQ27011, SQ9085, PB3552, SQ27066, SQ26977, and PB11300,it should be apparent to persons skilled in the art that other primersand/or probes can be designed to either amplify and/or hybridize tosequences within SEQ ID NO:1 which are unique to, and useful for,detecting the presence of DNA derived from event MON 87411 in a sample.

Based on molecular and sequence analysis, PCR assays for eventidentification assays were developed for event MON 87411. Followingstandard molecular biology laboratory practices, the parameters ofeither a standard PCR assay or a TaqMan® PCR assay were optimized witheach set of primer pairs and probes (i.e. probes labeled with afluorescent tag such as 6FAM™) used to detect the presence of DNAderived from event MON 87411 in a sample (SQ27011, SQ9085, and/orPB3552, or SQ27066, SQ26977, and/or PB11300). Generally, the parameterswhich were optimized included primer and probe concentration, amount oftemplate DNA, and PCR amplification cycling parameters. A control forthe PCR reaction included primers (S Q20221 (SEQ ID NO:38) and SQ20222(SEQ ID NO:40)) and/or probe (PB10065 (SEQ ID NO:39)) (probe labeledwith a fluorescent tag such as VIC™), which are specific for an internalcontrol, single copy gene in the corn genome. One of skill in the artwill know how to design other PCR primers specific for a single copygene in the corn genome which can be used to amplify an amplicon to beused as an internal control probe, or as an internal control in a PCRassay (e.g. TaqMan®). DNA was extracted from leaf tissue for each of thefollowing: [1] leaf sample to be analyzed; [2] negative control(non-transgenic corn DNA); [3] negative water control (no template); and[4] positive control MON 87411 DNA. Detection of the amplicons from astandard PCR assay would be visualization by DNA gel electrophoresis,and for a TaqMan® PCR assay by fluorescence detection.

A zygosity assay is useful for determining if a plant comprising anevent is homozygous for the event DNA; that is comprising the exogenousDNA in the same location on each chromosome of a chromosomal pair; orheterozygous for an event DNA, that is comprising the exogenous DNA ononly one chromosome of a chromosomal pair; or is null for the event DNA,that is wildtype. The zygosity of a corn plant containing event MON87411 can be determined by thermal amplification (PCR) or by endpointTaqMan® methods. For example, for PCR amplification, the primer pairSQ27011 (SEQ ID NO:18) and SQ26977 (SEQ ID NO:22) hybridize within thegenomic DNA flanking the event MON87411 insert. This primer pair willgenerate an amplicon which is 11323 nucleotides in length when DNAderived from event MON 87411 is present in the sample. This same primerpair will generate an amplicon which is only about 150 nucleotides longwhen corn DNA in the sample is not derived from event MON 87411. On DNAgel electrophoresis, a single band of 11323 bp is indicative that theDNA in the sample is from a homozygous MON 87411 event, a single band ofabout 150 bp is indicative that the DNA in the sample is not from a MON87411 event, and the presence of both a band of 11323 bp and a band ofabout 150 bp is indicative that the DNA in the sample is from a cornplant heterozygous for MON 87411 event.

A TaqMan® assay can be developed to determine the zygosity of a cornplant containing event MON 87411. For this assay, three or four primersand two probes would be designed where [1] a first primer pair and afirst probe are specific for detecting the presence of event MON 87411DNA in a sample, and [2] a second primer pair, different from the firstprimer pair, and a second probe, different from the first probe, arespecific for detecting the presence of wildtype corn DNA (i.e., samplenot containing event MON 87411). In a TaqMan®, or similar assay, afluorescent signal only from the first probe is indicative of anddiagnostic for a plant homozygous for event MON 87411; a fluorescentsignal from both the first probe and second probe is indicative of anddiagnostic for a plant heterozygous for event MON 87411; and afluorescent signal only from the second probe is indicative of anddiagnostic for a plant which is homozygous for the wildtype allele(i.e., is null for event MON 87411).

Example 5

This example describes the superior protection of plant comprising eventMON 87411 from corn rootworm damage when compared to current commercialproducts (MON 88017 and DAS-59122-7) and negative control plants.Efficacy field trials were conducted comparing 135 plants each of eventMON 87411, MON 88017, DAS-59122-7, and negative controls. Root damageratings (RDR) were collected, and the percentage plants with an RDR lessthan the economic injury level (0.25 RDR) is shown in Table 8.

Table 8 shows that only about 4% of plants containing event MON 87411exhibited RDRs greater than the economic threshold of 0.25 RDR. Incontrast, 22% of the commercially available plants containing MON 88017exhibited RDRs greater than the economic threshold of 0.25 RDR. And, 20%of the commercially available plants containing DAS-59122-7 exhibitedRDRs greater than the economic threshold of 0.25 RDR. And, 96% of thenegative control plants exhibited RDRs greater than the economicthreshold of 0.25 RDR. The conclusion from these data is that event MON87411 is clearly superior at providing protection from corn rootwormdamage as compared to commercial products MON 88071 and DAS-59122-7, anda negative control.

TABLE 8 Results of efficacy field trial with the approximate percentageof plants exhibiting ≤ 0.25 RDR. Approximate percentage of plantsexhibiting ≤ Event tested 0.25 RDR event MON 87411 96 MON 88017 78DAS-59122-7 80 negative control plants  4

Trial included 135 plants for each event tested.

Efficacy green house trials were conducted to test the performance ofevent MON 87411 with extreme infestation pressure of corn root worm. Inthis trial the following event were evaluated: event MON 87411, an eventfrom transformation with DNA vector #890 expressing only the dsRNA; MON88017; DAS-59122-7; and negative control. For these high-pressureefficacy trials, the corn plants under evaluation were grown in pots ina green house. Extreme infestation pressure was achieved by sequentialinfestation of each potted plant with approximately 2,000 WCR eggs perpot at their V2 growth stage, and, at 4 additional times occurring at 1to 1½ week intervals with approximately 1,000 WCR eggs per pot perinfestation for a total of approximately 6,000 WCR eggs added to eachpot. Plant roots were removed, washed, and rated for RDR at their VTgrowth stage. The roots from all thirteen (N=13) negative control plantsexhibited maximum root damage, or an absolute RDR of 3 RDR. Theseresults illustrate that event MON 87411 is more superior to other cornevents available for controlling corn rootworm (Table 9).

TABLE 9 Root Damage Rating (RDR) under high corn rootworm infestationpressure. Lower and Upper Average 95% confidence Event RDR limitsNegative Control 3.0 Absolute (N = 13) only dsRNA 0.36 0.17/0.54 (N =11) MON 88017 2.1 1.8/2.4 (N = 11) DAS-59122-7 0.29 0.17/0.42 (N = 16)MON 87411 0.06 0.03/0.08 (N = 13) (N = the number of plants evaluated).

One measure of efficacy of corn rootworm transgenic events is by adetermining the emergence of adult beetles from the potted soil ofplants cultivated in a green house. To determine adult corn rootwormbeetle emergence from the soil of event MON 87411 plants grown in pots,10 to 15 plants were germinated in pots containing soil infested withWCR eggs, similar to that described above. Throughout the growth period,each corn plant was covered with mesh bag to contain any emerging adultbeetles.

Counts of above ground adult beetles were made at 6, 12, and 18 weeksafter plant emergence, and at the end of the trial the roots wereevaluated for RDR. Plants containing event MON 87411 were compared tonegative control plants, and other corn rootworm protective transgenicevents. The results were that significantly fewer beetles were observedto emerge from soils in which event MON 87411 plants were pottedcompared to the other corn rootworm protective transgenic events,illustrating the superior properties of event MON 87411 to protectagainst corn rootworm damage.

Example 6

This example illustrates that the orientation of expression of twodifferent promoters in a corn cell, each driving expression of adifferent corn rootworm toxic agent, can result in significantlyimproved ratios of transgenic events exhibiting efficacy when providedin the diet of corn rootworm larvae.

Corn cells were transformed with one of four different planttransformation vectors, pMON120417, pMON120434, pMON120416, orpMON120419, and transgenic events were obtained that were regeneratedinto transgenic corn plants.

With reference to FIG. 4 , all of the plant transformation vectorscontain three expression cassettes 1, 2, and 3, bounded on one end by anAgrobacterium left border (LB), and at the opposite end by anAgrobacterium right border (RB). A corn rootworm toxic dsRNA isexpressed from cassette 1 in all four vectors from an enhancedCauliflower mosaic virus 35S (e35S) promoter. A corn rootworm toxinprotein, Cry3Bb, in vectors pMON120417, pMON120434 is expressed fromcassette 2 from a Zm.PIIG promoter. A corn rootworm toxin protein,Cry3Bb, in vectors pMON120416, pMON120419 is expressed from cassette 2from an Os.Rcc3 promoter. In all four vectors, a protein, conferringglyphosate herbicide tolerance, CTP-EPSPS CP4, is expressed fromcassette 3 from an Os.TubA3 promoter. In all four vectors cassette 1 andcassette 3 are in the same relative orientation. With reference to FIG.4 , the block arrows indicate the direction of expression from thepromoter in each of the respective cassettes.

The relative orientation of cassette 2 in vectors pMON120417 andpMON120434 is reversed, as illustrated by the block arrows (FIG. 4 )indicating the direction of expression from the promoter. Expression ofCry3Bb corn rootworm toxin protein in pMON120417 from cassette 2 isdivergent from the direction of expression of the corn rootworm toxicdsRNA expressed from cassette 1. Expression of Cry3Bb corn rootwormtoxin protein in pMON120434 from cassette 2 is in the same orientationas expression of the corn rootworm toxic dsRNA from cassette 1.

The relative orientation of cassette 2 in vectors pMON120416 andpMON120419 is reversed, as illustrated by the block arrows (FIG. 4 )indicating the direction of expression from the promoter. Expression ofCry3Bb corn rootworm toxin protein in pMON120416 from cassette 2 isdivergent from the direction of expression of the corn rootworm toxicdsRNA expressed from cassette 1. Expression of Cry3Bb corn rootwormtoxin protein in pMON120419 from cassette 2 is in the same orientationas expression of the corn rootworm toxic dsRNA from cassette 1.

As seen from Table 10, when tissue from transgenic corn plants wasprovided in the diet of Diabrotica species of corn root worm, the plantsgenerated by transformation with either construct pMON120417 orpMON120416 (divergent expression of the corn rootworm toxic components)was more efficacious with respect to pesticidal activity when comparedto plants generated by transformation with either construct pMON120434or pMON120419 (tandem or same orientation of expression) (Table 10). Theratio of efficacious events generated from transformation using thevectors pMON120417 and pMON120416, compared to the ratio of efficaciousevents from the vectors pMON120416 and pMON120419, was significantlygreater as shown by the data in Table 10. For example, for eventsgenerated from vector pMON120417 with the divergent promoter drivenexpression of the corn rootworm toxic components, 11 of 43 events, oralmost 25% of the events exhibited rootworm efficacious control. Incontrast, there were no efficacious events obtained for events generatedfrom vector pMON120434 with the promoter driven expression in the tandemorientation of the corn rootworm toxic components. For events generatedfrom vector pMON120416 with the divergent promoter driven expression ofthe corn rootworm toxic components, 17 of 27 events, or about 63% of theevents exhibited rootworm efficacious control. In contrast, there onlyabout 18.5% efficacious events obtained for events generated from vectorpMON120419 with the promoter driven expression in the tandem orientationof the corn rootworm toxic components. These data demonstrate thesignificantly improved number of efficacious events, and improved ratiosof transgenic events exhibiting efficacy, when transgenic corn plantsare generated from a plant transformation vector with two differentpromoters each driving expression in divergent directions of twodifferent corn rootworm toxic agents, and the transgenic corn plants areprovided in the diet of corn rootworm larvae.

TABLE 10 Results showing the number of R0 events and the numberefficacious events obtained from four plant transformation vectors. No.of R0 # Efficacious Construct Events events pMON120417 43 11 pMON1204348 0 pMON120416 27 17 pMON120419 43 8

Example 7

To produce corn plants or plant parts thereof which comprise enhancedagronomic, insecticidal, or herbicidal properties, corn plantscontaining event MON 87411 can be crossed with corn plants containingpotentially any other corn event or combination thereof and phenotypesevaluated to determine the resulting properties of the progeny plants.As a non-limiting example, MON 87411 can be crossed with corn plantsincluding one or more combinations, of the following: DAS-59122-7;MIR604; MON 89034; MON 87411; MON 87427; TC1507; 5307; DAS-06275-8;BT176; BT11; and MIR162.

What is claimed is:
 1. A method of detecting the presence of a DNAsegment diagnostic for corn event MON 87411 in a sample, the methodcomprising: (a) contacting said sample with a pair of DNA moleculescomprising a first DNA molecule and a second DNA molecule different fromthe first DNA molecule, wherein said first and second DNA molecules eachcomprise a polynucleotide segment of sufficient length of contiguousnucleotides of SEQ ID NO:1 or SEQ ID NO:2 or SEQ ID NO:3 to function asDNA primers when used together in an amplification reaction with asample containing corn event MON 87411 template DNA to produce anamplicon diagnostic for said corn event MON87411 DNA in said sample; (b)performing an amplification reaction sufficient to produce said DNAamplicon; and (c) detecting the presence of said DNA amplicon in saidreaction, wherein said DNA amplicon comprises the nucleotide sequenceselected from the group consisting of SEQ ID NO:2, SEQ ID NO:3, SEQ IDNO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10,SEQ ID NO:21, and SEQ ID NO:25, and wherein said detecting the presenceof said amplicon is diagnostic for the presence of corn event MON 87411DNA in said sample.
 2. The method of claim 1, wherein said samplecomprises a corn plant, corn plant cell, fractured corn cells, cornseed, corn plant part, corn plant tissue, or commodity corn product. 3.The method of claim 1, wherein detecting the presence of said DNAamplicon comprises detecting by direct sequencing, detectinghybridization of a DNA probe to said DNA amplicon, or detecting the sizeor composition of said DNA amplicon.
 4. The method of claim 1, whereinsaid first DNA molecule is comprised in the corn genomic DNA region ofSEQ ID NO:1 and said second DNA molecule is comprised in the insertedtransgenic DNA region of SEQ ID NO:1.
 5. The method of claim 1, whereinsaid pair of DNA molecules comprises the nucleotide sequence of SEQ IDNO:18 and SEQ ID NO:20, or SEQ ID NO:22 and SEQ ID NO:24.
 6. The methodof claim 3, wherein said detecting of the hybridization of said DNAprobe to said DNA amplicon comprises: (i) contacting said DNA ampliconwith said DNA probe; (ii) subjecting said DNA amplicon and said DNAprobe to stringent hybridization conditions; and (iii) detectinghybridization of said DNA probe to said amplicon, wherein detectionhybridization of said DNA probe to said amplicon is diagnostic for thepresence of said corn event.
 7. The method of claim 6, wherein said DNAprobe comprises at least one detectable label.
 8. The method of claim 7,wherein said detectable label is a florescent tag, radioactive tag,antibody based tag, or chemiluminescent tag.
 9. The method of claim 7,wherein detecting hybridization of said DNA probe to said ampliconcomprises detecting said detectable label.
 10. The method of claim 6,wherein the DNA probe comprises the nucleotide sequence of SEQ ID NO:19or SEQ ID NO:23.